Crystalline forms of a Bruton&#39;s tyrosine kinase inhibitor

ABSTRACT

Described herein is the Bruton&#39;s tyrosine kinase (Btk) inhibitor 1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one, including crystalline forms, solvates and pharmaceutically acceptable salts thereof. Also disclosed are pharmaceutical compositions that include the Btk inhibitor, as well as methods of using the Btk inhibitor, alone or in combination with other therapeutic agents, for the treatment of autoimmune diseases or conditions, heteroimmune diseases or conditions, cancer, including lymphoma, and inflammatory diseases or conditions.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/824,630, filed Mar. 19, 2020, which is a continuation of U.S.application Ser. No. 16/690,788, filed Nov. 21, 2019, now abandoned,which is a continuation of U.S. patent application Ser. No. 16/393,775,filed Apr. 24, 2019, now abandoned, which is a continuation of U.S.patent application Ser. No. 16/111,014, filed Aug. 23, 2018, now U.S.Pat. No. 10,294,232, issued May 21, 2019, which is a continuation ofU.S. patent application Ser. No. 15/900,660, filed Feb. 20, 2018, nowU.S. Pat. No. 10,106,548, issued Oct. 23, 2018, which is a continuationof U.S. patent application Ser. No. 15/887,744, filed Feb. 2, 2018, nowabandoned, which is a continuation of U.S. patent application Ser. No.15/645,319, filed Jul. 10, 2017, now abandoned, which is a continuationof U.S. patent application Ser. No. 15/497,896, filed Apr. 26, 2017, nowU.S. Pat. No. 9,725,455, issued Aug. 8, 2017, which is a continuation ofU.S. application Ser. No. 15/386,118, filed Dec. 21, 2016, now U.S. Pat.No. 9,713,617, issued Jul. 25, 2017, which is a divisional of U.S.patent application Ser. No. 14/405,317, filed Dec. 3, 2014, now U.S.Pat. No. 9,540,382, issued Jan. 10, 2017, which is the U.S. NationalStage application of PCT/US2013/043888, filed Jun. 3, 2013, which claimsthe benefit of U.S. Provisional Patent Application No. 61/655,381entitled “CRYSTALLINE FORMS OF A BRUTON'S TYROSINE KINASE INHIBITOR”filed Jun. 4, 2012, each of which is herein incorporated by reference inits entirety.

FIELD OF THE INVENTION

Described herein is the Bruton's tyrosine kinase (Btk) inhibitor1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one,including crystalline forms, solvates and pharmaceutically acceptablesalts thereof, as well as pharmaceutical compositions that include theBtk inhibitor and methods of using the Btk inhibitor in the treatment ofdiseases or conditions that would benefit from inhibition of Btkactivity.

BACKGROUND OF THE INVENTION

Bruton's tyrosine kinase (Btk), a member of the Tec family ofnon-receptor tyrosine kinases, is a key signaling enzyme expressed inall hematopoietic cells types except T lymphocytes and natural killercells. Btk plays an essential role in the B-cell signaling pathwaylinking cell surface B-cell receptor (BCR) stimulation to downstreamintracellular responses.

Btk is a key regulator of B-cell development, activation, signaling, andsurvival. In addition, Btk plays a role in a number of otherhematopoetic cell signaling pathways, e.g., Toll like receptor (TLR) andcytokine receptor-mediated TNF-α production in macrophages, IgE receptor(FcepsilonRI) signaling in Mast cells, inhibition of Fas/APO-1 apoptoticsignaling in B-lineage lymphoid cells, and collagen-stimulated plateletaggregation.

1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis also known by its IUPAC name as1-{(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl}prop-2-en-1-oneor 2-Propen-1-one,1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl-,and has been given the USAN name, ibrutinib. The various names given foribrutinib are used interchangeably herein.

SUMMARY OF THE INVENTION

Described herein is the Btk inhibitor1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one,including pharmaceutically acceptable solvates (including hydrates),polymorphs, and amorphous phases, and methods of uses thereof. Alsodescribed are pharmaceutically acceptable salts of the Btk inhibitor,including pharmaceutically acceptable solvates (including hydrates),polymorphs, and amorphous phases, and methods of uses thereof.1-((R)-3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one,as well as the pharmaceutically acceptable salts thereof, are used inthe manufacture of medicaments for the treatment of diseases orconditions that are associated with Btk activity.1-((R)-3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis an irreversible Btk inhibitor.

Also described herein are methods for preparing crystalline forms of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one.Further described are pharmaceutical compositions that include thecrystalline forms and methods of using the Btk inhibitor in thetreatment of diseases or conditions (including diseases or conditionswherein irreversible inhibition of Btk provides therapeutic benefit to amammal having the disease or condition).

In one embodiment is anhydrous1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one.

In another embodiment is crystalline anhydrous1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one.

In a further embodiment is amorphous anhydrous1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one.

In one aspect is a solvate of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one.

In one embodiment is a solvate, wherein1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis solvated with methyl isobutyl ketone (MIBK), toluene or methanol. Inone embodiment is a solvate, wherein1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis solvated with methyl isobutyl ketone (MIBK) or toluene. In oneembodiment is a solvate, wherein1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis solvated with methanol.

In a further embodiment, the solvate is anhydrous.

In another embodiment the solvate is crystalline.

In yet another embodiment the solvate is amorphous.

In one aspect, described herein is a crystalline Form A of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-onethat has at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 1;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 5.7±0.1° 2-Theta, 13.6±0.1° 2-Theta,        16.1±0.1° 2-Theta, 18.9±0.1° 2-Theta, 21.3±0.1° 2-Theta, and        21.6±0.1° 2-Theta;    -   (c) substantially the same X-ray powder diffraction (XRPD)        pattern post storage at 40° C. and 75% RH for at least a week;    -   (d) substantially the same X-ray powder diffraction (XRPD)        pattern post storage at 25° C. and 97% RH for at least a week;    -   (e) Infrared (IR) spectrum substantially similar to the one set        forth in FIG. 2;    -   (f) Infrared (IR) spectrum weak peaks at about 1584 cm⁻¹, about        1240 cm⁻¹, about 1147 cm⁻¹, about 1134 cm⁻¹, about 1099 cm⁻¹,        and about 1067 cm⁻¹;    -   (g) a DSC thermogram substantially similar to the one set forth        in FIG. 3;    -   (h) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 4;    -   (i) a DSC thermogram with an endotherm having an onset at about        154° C. and a peak at about 157° C. and an exotherm at about        159° C.;    -   (j) non-hygroscopicity;    -   (k) an observed aqueous solubility of about 0.013 mg/mL at about        pH 8;    -   or    -   (l) combinations thereof.

In some embodiments, the crystalline Form A has an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 1. Insome embodiments, the crystalline Form A has an X-ray powder diffraction(XRPD) pattern with characteristic peaks at 5.7±0.1° 2-Theta, 13.6±0.1°2-Theta, 16.1±0.1° 2-Theta, 18.9±0.1° 2-Theta, 21.3±0.1° 2-Theta, and21.6±0.1° 2-Theta. In some embodiments, the crystalline Form A hassubstantially the same X-ray powder diffraction (XRPD) pattern poststorage at 40° C. and 75% RH for at least a week. In some embodiments,the crystalline Form A has substantially the same X-ray powderdiffraction (XRPD) pattern post storage at 25° C. and 97% RH for atleast a week. In some embodiments, the crystalline Form A has anInfrared (IR) spectrum substantially similar to the one set forth inFIG. 2. In some embodiments, the crystalline Form A has an Infrared (IR)spectrum weak peaks at about 1584 cm⁻¹, about 1240 cm⁻¹, about 1147cm⁻¹, about 1134 cm⁻¹, about 1099 cm⁻¹, and about 1067 cm⁻¹. In someembodiments, the crystalline Form A has a melting temperature of about155-156° C. In some embodiments, the crystalline Form A has a DSCthermogram substantially similar to the one set forth in FIG. 3. In someembodiments, the crystalline Form A has a thermo-gravimetric analysis(TGA) thermogram substantially similar to the one set forth in FIG. 4.In some embodiments, the crystalline Form A has a DSC thermogram with anendotherm having an onset at about 154° C. and a peak at about 157° C.and an exotherm at about 159° C. In some embodiments, the crystallineForm A is non-hygroscopic. In some embodiments, the crystalline Form Ahas an observed aqueous solubility of about 0.013 mg/mL at about pH 8.In some embodiments, the crystalline Form A that is characterized ashaving properties (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), and(k). In some embodiments, the crystalline Form A was obtained from ethylacetate, isopropyl acetate, tetrahydrofuran, methyl isobutyl ketone(MIBK), methyl ethyl ketone (MEK), nitromethane, methanol, ethanol,acetonitrile, dioxane, methyl tert-butyl ether (MTBE), anisole, acetone,heptanes, a methanol/water mixture or an acetone/heptane mixture. Insome embodiments, the crystalline Form A was obtained from amethanol/water mixture. In some embodiments, the crystalline Form A isunsolvated. In some embodiments, the crystalline Form A is anhydrous.

In one aspect, described herein is a crystalline Form B of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-onethat has at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 5;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 5.2±0.1° 2-Theta, 10.2±0.1° 2-Theta,        16.5±0.1° 2-Theta, 18.5±0.1° 2-Theta, and 20.8±0.1° 2-Theta;    -   (c) substantially the same X-ray powder diffraction (XRPD)        pattern post storage at 40° C. and 75% RH for at least a week;    -   (d) substantially the same X-ray powder diffraction (XRPD)        pattern post storage at 25° C. and 97% RH for at least a week;    -   (e) Infrared (IR) spectrum substantially similar to the one set        forth in FIG. 6;    -   (f) Infrared (IR) spectrum weak peaks at about 1586 cm⁻¹, about        1573 cm⁻¹, about 1562 cm⁻¹, about 1229 cm⁻¹, about 1141 cm⁻¹,        about 1103 cm⁻¹, about 1056 cm⁻¹, and about 1033 cm⁻¹;    -   (g) a DSC thermogram substantially similar to the one set forth        in FIG. 7;    -   (h) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 8;    -   (i) a DSC thermogram with an endotherm having an onset at about        99-106° C. and a peak at about 115-118° C.;    -   (j) an observed aqueous solubility of about 0.0096 mg/mL at a pH        of about 7.42;    -   or    -   (k) combinations thereof.

In some embodiments, the crystalline Form B has an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 5. Insome embodiments, the crystalline Form B has an X-ray powder diffraction(XRPD) pattern with characteristic peaks at 5.2±0.1° 2-Theta, 10.2±0.1°2-Theta, 16.5±0.1° 2-Theta, 18.5±0.1° 2-Theta, and 20.8±0.1° 2-Theta. Insome embodiments, the crystalline Form B has an substantially the sameX-ray powder diffraction (XRPD) pattern post storage at 40° C. and 75%RH for at least a week. In some embodiments, the crystalline Form B hasan substantially the same X-ray powder diffraction (XRPD) pattern poststorage at 25° C. and 97% RH for at least a week. In some embodiments,the crystalline Form B has an Infrared (IR) spectrum substantiallysimilar to the one set forth in FIG. 6. In some embodiments, thecrystalline Form B has an Infrared (IR) spectrum weak peaks at about1586 cm⁻¹, about 1573 cm⁻¹, about 1562 cm⁻¹, about 1229 cm⁻¹, about 1141cm⁻¹, about 1103 cm⁻¹, about 1056 cm⁻¹, and about 1033 cm⁻¹. In someembodiments, the crystalline Form B has a DSC thermogram substantiallysimilar to the one set forth in FIG. 7. In some embodiments, thecrystalline Form B has a thermo-gravimetric analysis (TGA) thermogramsubstantially similar to the one set forth in FIG. 8. In someembodiments, the crystalline Form B has a DSC thermogram with anendotherm having an onset at about 99-106° C. and a peak at about115-118° C. In some embodiments, the crystalline Form B has an observedaqueous solubility of about 0.0096 mg/mL at a pH of about 7.42. In someembodiments, the crystalline Form B that is characterized as havingproperties (a), (b), (c), (d), (e), (f), (g), (h), (i), and (j). In someembodiments, the crystalline Form B was obtained from a mixture ofmethanol and water. In some embodiments, the crystalline Form B isunsolvated. In some embodiments, the crystalline Form B is anhydrous.

In one aspect, described herein is a crystalline Form C of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-onethat has at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 9;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 7.0±0.1° 2-Theta, 14.0±0.1° 2-Theta,        15.7±0.1° 2-Theta, 18.2±0.1° 2-Theta, 19.1±0.1° 2-Theta,        19.5±0.1° 2-Theta, 20.3±0.1° 2-Theta, 22.1±0.1° 2-Theta, and        22.9±0.1° 2-Theta;    -   (c) a DSC thermogram substantially similar to the one set forth        in FIG. 10;    -   (d) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 11;    -   (e) a DSC thermogram with an endotherm having an onset at about        134-135° C. and a peak at about 137-139° C.;    -   or    -   (f) combinations thereof.

In some embodiments, the crystalline Form C has an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 9. Insome embodiments, the crystalline Form C has an X-ray powder diffraction(XRPD) pattern with characteristic peaks at 7.0±0.1° 2-Theta, 14.0±0.1°2-Theta, 15.7±0.1° 2-Theta, 18.2±0.1° 2-Theta, 19.1±0.1° 2-Theta,19.5±0.1° 2-Theta, 20.3±0.1° 2-Theta, 22.1±0.1° 2-Theta, and 22.9±0.1°2-Theta. In some embodiments, the crystalline Form C has a DSCthermogram substantially similar to the one set forth in FIG. 10. Insome embodiments, the crystalline Form C has a thermo-gravimetricanalysis (TGA) thermogram substantially similar to the one set forth inFIG. 11. In some embodiments, the crystalline Form C has a DSCthermogram with an endotherm having an onset at about 134-135° C. and apeak at about 137-139° C. In some embodiments, the crystalline Form Cthat is characterized as having properties (a), (b), (c), (d), and (e).In some embodiments, the crystalline Form C was obtained from a mixtureof methanol and water. In some embodiments, the crystalline Form C isunsolvated. In some embodiments, the crystalline Form C is anhydrous.

In one aspect, described herein is a crystalline Form D of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-onethat has at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 12;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 7.2±0.1° 2-Theta, 8.0±0.1° 2-Theta,        9.2±0.1° 2-Theta, 14.5±0.1° 2-Theta, 18.5±0.1° 2-Theta,        19.5±0.1° 2-Theta, 20.7±0.1° 2-Theta, 21.0±0.1° 2-Theta,        21.9±0.1° 2-Theta, and 22.4±0.1° 2-Theta;    -   (c) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 13;    -   or    -   (d) combinations thereof.

In some embodiments, the crystalline Form D has an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 12.In some embodiments, the crystalline Form D has an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 7.2±0.1°2-Theta, 8.0±0.1° 2-Theta, 9.2±0.1° 2-Theta, 14.5±0.1° 2-Theta,18.5±0.1° 2-Theta, 19.5±0.1° 2-Theta, 20.7±0.1° 2-Theta, 21.0±0.1°2-Theta, 21.9±0.1° 2-Theta, and 22.4±0.1° 2-Theta. In some embodiments,the crystalline Form D has a thermo-gravimetric analysis (TGA)thermogram substantially similar to the one set forth in FIG. 13. Insome embodiments, the crystalline Form D that is characterized as havingproperties (a), (b), and (c). In some embodiments, the crystalline FormD was obtained from methyl isobutyl ketone (MIBK). In some embodiments,the crystalline Form D is solvated. In some embodiments, the crystallineForm D is solvated with methyl isobutyl ketone (MIBK).

In one aspect, described herein is a crystalline Form E of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-onethat has at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 14;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 7.8±0.1° 2-Theta, 8.8±0.1° 2-Theta,        16.1±0.1° 2-Theta, 18.1±0.1° 2-Theta, 19.3±0.1° 2-Theta,        19.5±0.1° 2-Theta, 20.5±0.1° 2-Theta, 21.6±0.1° 2-Theta, and        25.2±0.1° 2-Theta;    -   (c) a DSC thermogram substantially similar to the one set forth        in FIG. 15;    -   (d) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 15;    -   or    -   (e) combinations thereof.

In some embodiments, the crystalline Form E has an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 14.In some embodiments, the crystalline Form E has an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 7.8±0.1°2-Theta, 8.8±0.1° 2-Theta, 16.1±0.1° 2-Theta, 18.1±0.1° 2-Theta,19.3±0.1° 2-Theta, 19.5±0.1° 2-Theta, 20.5±0.1° 2-Theta, 21.6±0.1°2-Theta, and 25.2±0.1° 2-Theta. In some embodiments, the crystallineForm E has a DSC thermogram substantially similar to the one set forthin FIG. 15. In some embodiments, the crystalline Form E has athermo-gravimetric analysis (TGA) thermogram substantially similar tothe one set forth in FIG. 15. In some embodiments, the crystalline FormE that is characterized as having properties (a), (b), (c), and (d). Insome embodiments, the crystalline Form E was obtained from toluene. Insome embodiments, the crystalline Form E is solvated. In someembodiments, the crystalline Form E is solvated with toluene.

In one aspect, described herein is a crystalline Form F of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-onethat has at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 16;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 6.2±0.1° 2-Theta, 10.1±0.1° 2-Theta,        17.6±0.1° 2-Theta, 18.6±0.1° 2-Theta, 20.0±0.1° 2-Theta,        20.4±0.1° 2-Theta, 20.7±0.1° 2-Theta, 22.4±0.1° 2-Theta,        23.0±0.1° 2-Theta, 23.2±0.1° 2-Theta, 24.4±0.1° 2-Theta,        25.1±0.1° 2-Theta, 27.6±0.1° 2-Theta, 29.3±0.1° 2-Theta, and        29.7±0.1° 2-Theta;    -   (c) unit cell parameters substantially equal to the following at        100(2) K:

Crystal system Triclinic Space group P1 a 9.6332(3) Å α 105.762(3)° b9.7536(4) Å β  95.132(2)° c 15.0592(4) Å  γ 111.332(3)° V 1240.15(7) Å³Z 1 Density 1.308 Mg/m³ (calculated) Absorption 0.726 mm⁻¹ coefficientWavelength 1.54178 Å F(000) 518

-   -   or    -   (d) combinations thereof.

In some embodiments, crystalline Form F has an X-Ray powder diffraction(XRPD) pattern substantially the same as shown in FIG. 16. In someembodiments, crystalline Form F has an X-ray powder diffraction (XRPD)pattern with characteristic peaks at 6.2±0.1° 2-Theta, 10.1±0.1°2-Theta, 17.6±0.1° 2-Theta, 18.6±0.1° 2-Theta, 20.0±0.1° 2-Theta,20.4±0.1° 2-Theta, 20.7±0.1° 2-Theta, 22.4±0.1° 2-Theta, 23.0±0.1°2-Theta, 23.2±0.1° 2-Theta, 24.4±0.1° 2-Theta, 25.1±0.1° 2-Theta,27.6±0.1° 2-Theta, 29.3±0.1° 2-Theta, and 29.7±0.1° 2-Theta.

In some embodiments, crystalline Form F has unit cell parameterssubstantially equal to the following at 100(2) K:

Crystal system Triclinic Space group P1 a 9.6332(3) Å α 105.762(3)° b9.7536(4) Å β  95.132(2)° c 15.0592(4) Å  γ 111.332(3)° V 1240.15(7) Å³Z 1 Density 1.308 Mg/m³ (calculated) Absorption 0.726 mm⁻¹ coefficientWavelength 1.54178 Å F(000) 518

In some embodiments, crystalline Form F was obtained from methanol.

In some embodiments, crystalline Form F is solvated. In someembodiments, crystalline Form F is solvated with methanol.

In one aspect, described herein is a pharmaceutically acceptable salt of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one,wherein the pharmaceutically acceptable salt is an acid addition salt.In some embodiments, the pharmaceutically acceptable salt is amorphous.In some embodiments, the pharmaceutically acceptable salt iscrystalline.

In a further aspect are provided pharmaceutical compositions, whichinclude1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneas described herein, and at least one additional ingredient selectedfrom pharmaceutically acceptable carriers, diluents and excipients. Insome embodiments, the pharmaceutical composition comprises Form A. Insome embodiments, the pharmaceutical composition comprises Form B. Insome embodiments, the pharmaceutical composition comprises Form C. Insome embodiments, the pharmaceutical composition comprises Form D. Insome embodiments, the pharmaceutical composition comprises Form E. Insome embodiments, the pharmaceutical composition comprises Form F. Insome embodiments, the pharmaceutical composition is in a form suitablefor oral administration to a mammal. In some embodiments, thepharmaceutical composition is an oral solid dosage form. In someembodiments, the pharmaceutical composition comprises about 0.5 mg toabout 1000 mg of crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one.

In another aspect, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 mgs to about 200 mgs of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of a diluent;

(c) about 3 wt % to about 10 wt % of a disintegrating agent;

(d) about 2 wt % to about 7 wt % of a surfactant; and

(e) about 0.2 wt % to about 1.0 wt % of a lubricant.

In some embodiments, the diluent is selected from the group consistingof lactose, sucrose, dextrose, dextrates, maltodextrin, mannitol,xylitol, sorbitol, cyclodextrins, calcium phosphate, calcium sulfate,starches, modified starches, microcrystalline cellulose, microcellulose,and talc. In some embodiments the diluent is microcrystalline cellulose.In some embodiments, the disintegrating agent is selected from the groupconsisting of natural starch, a pregelatinized starch, a sodium starch,methylcrystalline cellulose, methylcellulose, croscarmellose,croscarmellose sodium, cross-linked sodium carboxymethylcellulose,cross-linked carboxymethylcellulose, cross-linked croscarmellose,cross-linked starch such as sodium starch glycolate, cross-linkedpolymer such as crospovidone, cross-linked polyvinylpyrrolidone, sodiumalginate, a clay, or a gum. In some embodiments, the disintegratingagent is croscarmellose sodium. In some embodiments, the surfactant isselected from the group consisting of sodium lauryl sulfate, sorbitanmonooleate, polyoxyethylene sorbitan monooleate, polysorbates,polaxomers, bile salts, glyceryl monostearate, copolymers of ethyleneoxide and propylene oxide. In some embodiments, the surfactant is sodiumlauryl sulfate. In some embodiments, the lubricant is selected from thegroup consisting of stearic acid, calcium hydroxide, talc, corn starch,sodium stearyl fumerate, stearic acid, sodium stearates, magnesiumstearate, zinc stearate, and waxes. In some embodiments, the lubricantis magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 mgs to about 200 mgs of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of microcrystalline cellulose;

(c) about 3 wt % to about 10 wt % of croscarmellose sodium;

(d) about 2 wt % to about 7 wt % of sodium lauryl sulfate; and

(e) about 0.2 wt % to about 1.0 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 wt % to about 50 wt % of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of microcrystalline cellulose;

(c) about 3 wt % to about 10 wt % of croscarmellose sodium;

(d) about 2 wt % to about 7 wt % of sodium lauryl sulfate; and

(e) about 0.2 wt % to about 1.0 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) 140 mgs of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) 45.9 wt % of microcrystalline cellulose;

(c) 7.0 wt % of croscarmellose sodium;

(d) 4.2 wt % of sodium lauryl sulfate; and

(e) 0.5 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) 140 mgs of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) 151.4 mgs of microcrystalline cellulose;

(c) 23.0 mgs of croscarmellose sodium;

(d) 14.0 mgs of sodium lauryl sulfate; and

(e) 1.6 mgs of magnesium stearate.

In another aspect, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 mgs to about 200 mgs of crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of a diluent;

(c) about 3 wt % to about 10 wt % of a disintegrating agent;

(d) about 2 wt % to about 7 wt % of a surfactant; and

(e) about 0.2 wt % to about 1.0 wt % of a lubricant.

In some embodiments, the diluent is selected from the group consistingof lactose, sucrose, dextrose, dextrates, maltodextrin, mannitol,xylitol, sorbitol, cyclodextrins, calcium phosphate, calcium sulfate,starches, modified starches, microcrystalline cellulose, microcellulose,and talc. In some embodiments the diluent is microcrystalline cellulose.In some embodiments, the disintegrating agent is selected from the groupconsisting of natural starch, a pregelatinized starch, a sodium starch,methylcrystalline cellulose, methylcellulose, croscarmellose,croscarmellose sodium, cross-linked sodium carboxymethylcellulose,cross-linked carboxymethylcellulose, cross-linked croscarmellose,cross-linked starch such as sodium starch glycolate, cross-linkedpolymer such as crospovidone, cross-linked polyvinylpyrrolidone, sodiumalginate, a clay, or a gum. In some embodiments, the disintegratingagent is croscarmellose sodium. In some embodiments, the surfactant isselected from the group consisting of sodium lauryl sulfate, sorbitanmonooleate, polyoxyethylene sorbitan monooleate, polysorbates,polaxomers, bile salts, glyceryl monostearate, copolymers of ethyleneoxide and propylene oxide. In some embodiments, the surfactant is sodiumlauryl sulfate. In some embodiments, the lubricant is selected from thegroup consisting of stearic acid, calcium hydroxide, talc, corn starch,sodium stearyl fumerate, stearic acid, sodium stearates, magnesiumstearate, zinc stearate, and waxes. In some embodiments, the lubricantis magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 mgs to about 200 mgs of crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of microcrystalline cellulose;

(c) about 3 wt % to about 10 wt % of croscarmellose sodium;

(d) about 2 wt % to about 7 wt % of sodium lauryl sulfate; and

(e) about 0.2 wt % to about 1.0 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 wt % to about 50 wt % of crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of microcrystalline cellulose;

(c) about 3 wt % to about 10 wt % of croscarmellose sodium;

(d) about 2 wt % to about 7 wt % of sodium lauryl sulfate; and

(e) about 0.2 wt % to about 1.0 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) 140 mgs of crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) 45.9 wt % of microcrystalline cellulose;

(c) 7.0 wt % of croscarmellose sodium;

(d) 4.2 wt % of sodium lauryl sulfate; and

(e) 0.5 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) 140 mgs of crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) 151.4 mgs of microcrystalline cellulose;

(c) 23.0 mgs of croscarmellose sodium;

(d) 14.0 mgs of sodium lauryl sulfate; and

(e) 1.6 mgs of magnesium stearate.

In some embodiments of the aforementioned pharmaceutical formulationembodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis crystalline Form A. In some embodiments of the aforementionedpharmaceutical formulation embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis crystalline Form B. In some embodiments of the aforementionedpharmaceutical formulation embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis crystalline Form C. In some embodiments of the aforementionedpharmaceutical formulation embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis crystalline Form D. In some embodiments of the aforementionedpharmaceutical formulation embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis crystalline Form E. In some embodiments of the aforementionedpharmaceutical formulation embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis crystalline Form F. In some embodiments of the aforementionedpharmaceutical formulation embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis a mixture of two or more crystalline forms selected from the groupconsisting of Form A, Form B, Form C, Form D, Form E, and Form F. Inanother embodiment of the aforementioned pharmaceutical formulationembodiments provided herein, is a pharmaceutical formulation wherein thedosage form is a hard gelatin capsule.

In another aspect, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 mgs to about 200 mgs of crystalline Form A of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of a diluent;

(c) about 3 wt % to about 10 wt % of a disintegrating agent;

(d) about 2 wt % to about 7 wt % of a surfactant; and

(e) about 0.2 wt % to about 1.0 wt % of a lubricant.

In some embodiments, the diluent is selected from the group consistingof lactose, sucrose, dextrose, dextrates, maltodextrin, mannitol,xylitol, sorbitol, cyclodextrins, calcium phosphate, calcium sulfate,starches, modified starches, microcrystalline cellulose, microcellulose,and talc. In some embodiments the diluent is microcrystalline cellulose.In some embodiments, the disintegrating agent is selected from the groupconsisting of natural starch, a pregelatinized starch, a sodium starch,methylcrystalline cellulose, methylcellulose, croscarmellose,croscarmellose sodium, cross-linked sodium carboxymethylcellulose,cross-linked carboxymethylcellulose, cross-linked croscarmellose,cross-linked starch such as sodium starch glycolate, cross-linkedpolymer such as crospovidone, cross-linked polyvinylpyrrolidone, sodiumalginate, a clay, or a gum. In some embodiments, the disintegratingagent is croscarmellose sodium. In some embodiments, the surfactant isselected from the group consisting of sodium lauryl sulfate, sorbitanmonooleate, polyoxyethylene sorbitan monooleate, polysorbates,polaxomers, bile salts, glyceryl monostearate, copolymers of ethyleneoxide and propylene oxide. In some embodiments, the surfactant is sodiumlauryl sulfate. In some embodiments, the lubricant is selected from thegroup consisting of stearic acid, calcium hydroxide, talc, corn starch,sodium stearyl fumerate, stearic acid, sodium stearates, magnesiumstearate, zinc stearate, and waxes. In some embodiments, the lubricantis magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 mgs to about 200 mgs of crystalline Form A of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of microcrystalline cellulose;

(c) about 3 wt % to about 10 wt % of croscarmellose sodium;

(d) about 2 wt % to about 7 wt % of sodium lauryl sulfate; and

(e) about 0.2 wt % to about 1.0 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) about 40 wt % to about 50 wt % of crystalline Form A of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) about 40 wt % to about 50 wt % of microcrystalline cellulose;

(c) about 3 wt % to about 10 wt % of croscarmellose sodium;

(d) about 2 wt % to about 7 wt % of sodium lauryl sulfate; and

(e) about 0.2 wt % to about 1.0 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) 140 mgs of crystalline Form A of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) 45.9 wt % of microcrystalline cellulose;

(c) 7.0 wt % of croscarmellose sodium;

(d) 4.2 wt % of sodium lauryl sulfate; and

(e) 0.5 wt % of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical formulation fororal administration comprising:

(a) 140 mgs of crystalline Form A of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

(b) 151.4 mgs of microcrystalline cellulose;

(c) 23.0 mgs of croscarmellose sodium;

(d) 14.0 mgs of sodium lauryl sulfate; and

(e) 1.6 mgs of magnesium stearate.

In another aspect provided herein, is a pharmaceutical formulationcomprising: a) about 40 mgs to about 200 mgs of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;b) about 40 wt % to about 50 wt % of a diluent; c) about 3 wt % to about10 wt % of a disintegrating agent; d) about 2 wt % to about 7 wt % of asurfactant; and e) about 0.2 wt % to about 1.0 wt % of a lubricant;wherein the formulation is in a unit dosage form in a blister pack, andsaid blister pack comprises metal or plastic foil. In some embodiments,is a pharmaceutical formulation comprising: a) 140 mgs of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;b) 45.9 wt % of microcrystalline cellulose; c) 7.0 wt % ofcroscarmellose sodium; d) 4.2 wt % of sodium lauryl sulfate; and e) 0.5wt % of magnesium stearate wherein the formulation is in a unit dosageform in a blister pack, and said blister pack comprises metal or plasticfoil.

In another embodiment is a package comprising one or more discreteblister pockets, wherein each blister pocket comprises a unit dosageform comprising:

a) about 40 mgs to about 200 mgs of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

b) about 40 wt % to about 50 wt % of a diluent; c) about 3 wt % to about10 wt % of a disintegrating agent; d) about 2 wt % to about 7 wt % of asurfactant; and

e) about 0.2 wt % to about 1.0 wt % of a lubricant;

wherein each blister pocket comprises metal or plastic foil.

In another aspect provided herein, is a pharmaceutical formulationcomprising: a) about 40 mgs to about 200 mgs of crystalline Form A of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;b) about 40 wt % to about 50 wt % of a diluent; c) about 3 wt % to about10 wt % of a disintegrating agent; d) about 2 wt % to about 7 wt % of asurfactant; and e) about 0.2 wt % to about 1.0 wt % of a lubricant;wherein the formulation is in a unit dosage form in a blister pack, andsaid blister pack comprises metal or plastic foil. In some embodiments,is a pharmaceutical formulation comprising: a) 140 mgs of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;b) 45.9 wt % of microcrystalline cellulose; c) 7.0 wt % ofcroscarmellose sodium; d) 4.2 wt % of sodium lauryl sulfate; and e) 0.5wt % of magnesium stearate wherein the formulation is in a unit dosageform in a blister pack, and said blister pack comprises metal or plasticfoil.

In another embodiment is a package comprising one or more discreteblister pockets, wherein each blister pocket comprises a unit dosageform comprising:

a) about 40 mgs to about 200 mgs of crystalline Form A of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one;

b) about 40 wt % to about 50 wt % of a diluent; c) about 3 wt % to about10 wt % of a disintegrating agent; d) about 2 wt % to about 7 wt % of asurfactant; and

e) about 0.2 wt % to about 1.0 wt % of a lubricant;

wherein each blister pocket comprises metal or plastic foil.

In one embodiment, a kit is provided which contains a multiplicity oforal dosage forms, such as tablets or capsules, packaging such as a jarcontaining the oral dosage forms, and instructions for use to administerthe oral dosage forms in accordance with the method described herein.Unit dose packaging such as blister packs provide a useful way ofpackaging the oral dosage form of the formulations described herein, andin other embodiments embody a kit when combined with instructions foruse. In other embodiments, detailed product information are includedwith the instructions for use in the kit. Blister packaging isparticularly useful with solid oral dosage forms and are in furtherembodiments useful for alternate day dosing schedules for example. Inone embodiment, solid unit dosage forms of the formulations describedherein included in a blister pack with instructions to administer one ormore tablets or capsules on a daily basis so that the dosage of theformulations described herein are sufficiently administered. In anotherembodiment, solid unit dosage forms are included in a blister pack withinstructions to administer one or more tablets or capsules on analternate day basis so that the dosage per day is sufficientlyadministered.

In one aspect, provided herein are methods for treating a patient byadministering Compound 1. In some embodiments, provided herein is amethod of inhibiting the activity of tyrsoine kinase(s), such as Btk, orof treating a disease, disorder, or condition, which would benefit frominhibition of tyrosine kinase(s), such as Btk, in a mammal, whichincludes administering to the mammal a therapeutically effective amountof Compound 1, or pharmaceutically acceptable salt, pharmaceuticallyactive metabolite, pharmaceutically acceptable prodrug, orpharmaceutically acceptable solvate.

In another aspect, provided herein is the use of Compound 1 forinhibiting Bruton's tyrosine kinase (Btk) activity or for the treatmentof a disease, disorder, or condition, which would benefit frominhibition of Bruton's tyrosine kinase (Btk) activity.

In some embodiments, crystalline Compound 1 is administered to a human.

In some embodiments, crystalline Compound 1 is orally administered.

In other embodiments, crystalline Compound 1 is used for the formulationof a medicament for the inhibition of tyrosine kinase activity. In someother embodiments, crystalline Compound 1 is used for the formulation ofa medicament for the inhibition of Bruton's tyrosine kinase (Btk)activity.

In one aspect, provided herein is a method of treating cancer in amammal comprising administering to the mammal a pharmaceuticalcomposition described herein comprising Compound 1. In some embodiments,the cancer is a B cell malignancy. In some embodiments, the cancer is aB cell malignancy selected from chronic lymphocytic leukemia (CLL)/smalllymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), diffuse large BCell lymphoma (DLBCL), and multiple myeloma. In some embodiments, thecancer is a lymphoma, leukemia or a solid tumor. In some embodiments,the cancer is diffuse large B cell lymphoma, follicular lymphoma,chronic lymphocytic lymphoma, chronic lymphocytic leukemia, B-cellprolymphocytic leukemia, lymphoplasmacytic lymphoma/Waldenstrammacroglobulinemia, splenic marginal zone lymphoma, plasma cell myeloma,plasmacytoma, extranodal marginal zone B cell lymphoma, nodal marginalzone B cell lymphoma, mantle cell lymphoma, mediastinal (thymic) large Bcell lymphoma, intravascular large B cell lymphoma, primary effusionlymphoma, burkitt lymphoma/leukemia, or lymphomatoid granulomatosis. Insome embodiments, where the subject is suffering from a cancer, ananti-cancer agent is administered to the subject in addition to one ofthe above-mentioned compounds. In one embodiment, the anti-cancer agentis an inhibitor of mitogen-activated protein kinase signaling

In one aspect, provided herein is a method of treating an inflammatoryor an autoimmune disease in a mammal comprising administering to themammal a pharmaceutical composition described herein comprisingCompound 1. In some embodiments, the inflammatory disease is asthma,appendicitis, blepharitis, bronchiolitis, bronchitis, bursitis,cervicitis, cholangitis, cholecystitis, colitis, conjunctivitis,cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis,endocarditis, endometritis, enteritis, enterocolitis, epicondylitis,epididymitis, fasciitis, fibrositis, gastritis, gastroenteritis,hepatitis, hidradenitis suppurativa, laryngitis, mastitis, meningitis,myelitis myocarditis, myositis, nephritis, oophoritis, orchitis,osteitis, otitis, pancreatitis, parotitis, pericarditis, peritonitis,pharyngitis, pleuritis, phlebitis, pneumonitis, pneumonia, proctitis,prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis,stomatitis, synovitis, tendonitis, tonsillitis, uveitis, vaginitis,vasculitis, or vulvitis. In some embodiments, the autoimmune disease isinflammatory bowel disease, arthritis, lupus, rheumatoid arthritis,psoriatic arthritis, osteoarthritis, Still's disease, juvenilearthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Ord'sthyroiditis, Graves' disease Sjagren's syndrome, multiple sclerosis,Guillain-Barré syndrome, acute disseminated encephalomyelitis, Addison'sdisease, opsoclonus-myoclonus syndrome, ankylosing spondylitisis,antiphospholipid antibody syndrome, aplastic anemia, autoimmunehepatitis, coeliac disease, Goodpasture's syndrome, idiopathicthrombocytopenic purpura, optic neuritis, scleroderma, primary biliarycirrhosis, Reiter's syndrome, Takayasu's arteritis, temporal arteritis,warm autoimmune hemolytic anemia, Wegener's granulomatosis, psoriasis,alopecia universalis, Behget's disease, chronic fatigue, dysautonomia,endometriosis, interstitial cystitis, neuromyotonia, scleroderma, orvulvodynia

Articles of manufacture including packaging material, Compound 1 withinthe packaging material, and a label that indicates that Compound 1 isused for inhibiting the activity of tyrosine kinase(s), such as Btk, areprovided.

In a further aspect, provided herein is a method of treating anautoimmune disease in a mammal, comprising administering Compound 1 tothe mammal.

In a further aspect, provided herein is a method of treating aheteroimmune disease or condition in a mammal, comprising administeringCompound 1 to the mammal.

In a further aspect, provided herein is a method of treating aninflammatory disease in a mammal, comprising administering Compound 1 tothe mammal.

In a further aspect, provided herein is a method of treating cancer in amammal, comprising administering Compound 1 to the mammal.

In a further aspect, provided herein is a method of treating athromboembolic disorder in a mammal, comprising administering Compound 1to the mammal. Thromboembolic disorders include, but are not limited to,myocardial infarct, angina pectoris, reocclusion after angioplasty,restenosis after angioplasty, reocclusion after aortocoronary bypass,restenosis after aortocoronary bypass, stroke, transitory ischemia, aperipheral arterial occlusive disorder, pulmonary embolism, or deepvenous thrombosis.

In another aspect are methods for modulating, including irreversiblyinhibiting the activity of Btk or other tyrosine kinases, wherein theother tyrosine kinases share homology with Btk by having a cysteineresidue (including a Cys 481 residue) that can form a covalent bond withCompound 1, in a mammal comprising administering to the mammal at leastonce an effective amount of Compound 1. In another aspect are methodsfor modulating, including including irreversibly inhibiting, theactivity of Btk in a mammal comprising administering to the mammal atleast once an effective amount of Compound 1. In another aspect aremethods for treating Btk-dependent or Btk mediated conditions ordiseases, comprising administering to the mammal at least once aneffective amount of Compound 1.

In another aspect are methods for treating inflammation comprisingadministering to the mammal at least once an effective amount ofCompound 1.

A further aspect are methods for the treatment of cancer comprisingadministering to the mammal at least once an effective amount ofCompound 1. The type of cancer may include, but is not limited to,pancreatic cancer and other solid or hematological tumors.

In another aspect are methods for treating respiratory diseasescomprising administering to the mammal at least once an effective amountof Compound 1. In a further embodiment of this aspect, the respiratorydisease is asthma. In a further embodiment of this aspect, therespiratory disease includes, but is not limited to, adult respiratorydistress syndrome and allergic (extrinsic) asthma, non-allergic(intrinsic) asthma, acute severe asthma, chronic asthma, clinicalasthma, nocturnal asthma, allergen-induced asthma, aspirin-sensitiveasthma, exercise-induced asthma, isocapnic hyperventilation, child-onsetasthma, adult-onset asthma, cough-variant asthma, occupational asthma,steroid-resistant asthma, seasonal asthma.

In another aspect are methods for preventing rheumatoid arthritis and/orosteoarthritis comprising administering to the mammal at least once aneffective amount of Compound 1.

In another aspect are methods for treating inflammatory responses of theskin comprising administering to the mammal at least once an effectiveamount of Compound 1. Such inflammatory responses of the skin include,by way of example, dermatitis, contact dermatitis, eczema, urticaria,rosacea, and scarring. In another aspect are methods for reducingpsoriatic lesions in the skin, joints, or other tissues or organs,comprising administering to the mammal an effective amount of Compound1.

In another aspect is the use of Compound 1 in the manufacture of amedicament for treating an inflammatory disease or condition in ananimal in which the activity of Btk or other tyrosine kinases, whereinthe other tyrosine kinases share homology with Btk by having a cysteineresidue (including a Cys 481 residue) that can form a covalent bond withat least one irreversible inhibitor described herein, contributes to thepathology and/or symptoms of the disease or condition. In one embodimentof this aspect, the tyrosine kinase protein is Btk. In another orfurther embodiment of this aspect, the inflammatory disease orconditions are respiratory, cardiovascular, or proliferative diseases.

In any of the aforementioned aspects are further embodiments in whichCompound 1 is (a) systemically administered to the mammal; (b)administered orally to the mammal; (c) intravenously administered to themammal; (d) administered by inhalation; (e) administered by nasaladministration; or (f) administered by injection to the mammal; (g)administered topically (dermal) to the mammal; (h) administered byophthalmic administration; or (i) administered rectally to the mammal.

In any of the aforementioned aspects are further embodiments comprisingsingle administration of Compound 1, including further embodiments inwhich Compound 1 is administered (i) once; (ii) multiple times over thespan of one day; (iii) continually; or (iv) continuously.

In any of the aforementioned aspects are further embodiments comprisingmultiple administrations of Compound 1, including further embodiments inwhich (i) Compound 1 is administered in a single dose; (ii) the timebetween multiple administrations is every 6 hours; (iii) Compound 1 isadministered to the mammal every 8 hours. In further or alternativeembodiments, the method comprises a drug holiday, wherein theadministration of Compound 1 is temporarily suspended or the dose ofCompound 1 being administered is temporarily reduced; at the end of thedrug holiday, dosing of Compound 1 is resumed. The length of the drugholiday can vary from 2 days to 1 year.

In some embodiments, in any of the embodiments disclosed herein(including methods, uses, formulations, combination therapy, etc.),Compound 1, or a pharmaceutically acceptable salt or solvate thereof,is: optically pure (i.e. greater than 99% chiral purity by HPLC). Insome embodiments, in any of the embodiments disclosed herein (includingmethods, uses, formulations, combination therapy, etc.), Compound 1, ora pharmaceutically acceptable salt or solvate thereof, is replaced with:a) Compound 1, or a pharmaceutically acceptable salt or solvate thereof,of lower chiral purity; b)1-((S)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one,or a pharmaceutically acceptable salt or solvate thereof of any opticalpurity; or c) racemic1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one,or a pharmaceutically acceptable salt or solvate thereof.

In any of the embodiments disclosed herein (including methods, uses,formulations, combination therapy, etc.), amorphous Compound 1 is used.In any of the embodiments disclosed herein (including methods, uses,formulations, combination therapy, etc.), crystalline Compound 1 isused. In any of the embodiments disclosed herein (including methods,uses, formulations, combination therapy, etc.), crystalline Compound 1(Form A) is used. In any of the embodiments disclosed herein (includingmethods, uses, formulations, combination therapy, etc.), crystallineCompound 1 (Form B) is used. In any of the embodiments disclosed herein(including methods, uses, formulations, combination therapy, etc.),crystalline Compound 1 (Form C) is used. In any of the embodimentsdisclosed herein (including methods, uses, formulations, combinationtherapy, etc.), crystalline Compound 1 (Form D) is used. In any of theembodiments disclosed herein (including methods, uses, formulations,combination therapy, etc.), crystalline Compound 1 (Form E) is used. Inany of the embodiments disclosed herein (including methods, uses,formulations, combination therapy, etc.), crystalline Compound 1 (FormF) is used.

In some embodiments, in any of the embodiments disclosed herein(including methods, uses, formulations, combination therapy, etc.),Compound 1, or a pharmaceutically acceptable salt thereof, is replacedwith an active metabolite of Compound 1. In some embodiments, the activemetabolite is in a crystalline form. In some embodiments, the activemetabolite is in an amorphous phase. In further embodiments themetabolite is isolated. In some embodiments, in any of the embodimentsdisclosed herein (including methods, uses, formulations, combinationtherapy, etc.), Compound 1, or a pharmaceutically acceptable saltthereof, is replaced with a prodrug of Compound 1, or a deuteratedanalog of Compound 1, or a pharmaceutically acceptable salt thereof.

Other objects, features and advantages of the methods and compositionsdescribed herein will become apparent from the following detaileddescription. It should be understood, however, that the detaileddescription and the specific examples, while indicating specificembodiments, are given by way of illustration only, since variouschanges and modifications within the spirit and scope of the presentdisclosure will become apparent to those skilled in the art from thisdetailed description. The section headings used herein are fororganizational purposes only and are not to be construed as limiting thesubject matter described. All documents, or portions of documents, citedin the application including, but not limited to, patents, patentapplications, articles, books, manuals, and treatises are herebyexpressly incorporated by reference in their entirety for any purpose.

INCORPORATION BY REFERENCE

All publications and patent applications mentioned in this specificationare herein incorporated by reference to the extent applicable andrelevant.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Illustrates an X-Ray powder diffraction (XRPD) pattern of FormA.

FIG. 2. Illustrates an Infrared (IR) spectrum of Form A.

FIG. 3. Illustrates DSC thermograms of Form A.

FIG. 4. Illustrates a thermo-gravimetric analysis (TGA) thermogram ofForm A.

FIG. 5. Illustrates an X-Ray powder diffraction (XRPD) pattern of FormB.

FIG. 6. Illustrates an Infrared (IR) spectrum of Form B.

FIG. 7. Illustrates DSC thermograms of Form B.

FIG. 8. Illustrates a thermo-gravimetric analysis (TGA) thermogram ofForm B.

FIG. 9. Illustrates an X-Ray powder diffraction (XRPD) pattern of FormC.

FIG. 10. Illustrates DSC thermograms of Form C.

FIG. 11. Illustrates a thermo-gravimetric analysis (TGA) thermogram ofForm C.

FIG. 12. Illustrates an X-Ray powder diffraction (XRPD) pattern of FormD.

FIG. 13. Illustrates a thermo-gravimetric analysis (TGA) thermogram ofForm D.

FIG. 14. Illustrates an X-Ray powder diffraction (XRPD) pattern of FormE.

FIG. 15. Illustrates a DSC thermogram and a thermo-gravimetric analysis(TGA) thermogram of Form E.

FIG. 16. Illustrates a simulated X-Ray powder diffraction (XRPD) patternof Form F.

DETAILED DESCRIPTION OF THE INVENTION

The diverse roles played by Btk signaling in various hematopoietic cellfunctions, e.g., B-cell receptor activation, suggests that smallmolecule Btk inhibitors, such as Compound 1, are useful for reducing therisk of or treating a variety of diseases affected by or affecting manycell types of the hematopoetic lineage including, e.g., autoimmunediseases, heteroimmune conditions or diseases, inflammatory diseases,cancer (e.g., B-cell proliferative disorders), and thromboembolicdisorders. Further, irreversible Btk inhibitor compounds, such asCompound 1, can be used to inhibit a small subset of other tyrosinekinases that share homology with Btk by having a cysteine residue(including a Cys 481 residue) that can form a covalent bond with theirreversible inhibitor.

In some embodiments, Compound 1 can be used in the treatment of anautoimmune disease in a mammal, which includes, but is not limited to,rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still'sdisease, juvenile arthritis, lupus, diabetes, myasthenia gravis,Hashimoto's thyroiditis, Ord's thyroiditis, Graves' disease Sjagren'ssyndrome, multiple sclerosis, Guillain-Barré syndrome, acutedisseminated encephalomyelitis, Addison's disease, opsoclonus-myoclonussyndrome, ankylosing spondylitisis, antiphospholipid antibody syndrome,aplastic anemia, autoimmune hepatitis, coeliac disease, Goodpasture'ssyndrome, idiopathic thrombocytopenic purpura, optic neuritis,scleroderma, primary biliary cirrhosis, Reiter's syndrome, Takayasu'sarteritis, temporal arteritis, warm autoimmune hemolytic anemia,Wegener's granulomatosis, psoriasis, alopecia universalis, Behget'sdisease, chronic fatigue, dysautonomia, endometriosis, interstitialcystitis, neuromyotonia, scleroderma, and vulvodynia.

In some embodiments, Compound 1 can be used in the treatment of aheteroimmune disease or condition in a mammal, which include, but arenot limited to graft versus host disease, transplantation, transfusion,anaphylaxis, allergies (e.g., allergies to plant pollens, latex, drugs,foods, insect poisons, animal hair, animal dander, dust mites, orcockroach calyx), type I hypersensitivity, allergic conjunctivitis,allergic rhinitis, and atopic dermatitis.

In some embodiments, Compound 1 can be used in the treatment of aninflammatory disease in a mammal, which includes, but is not limited toasthma, inflammatory bowel disease, appendicitis, blepharitis,bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis,cholecystitis, colitis, conjunctivitis, cystitis, dacryoadenitis,dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis,enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis,fibrositis, gastritis, gastroenteritis, hepatitis, hidradenitissuppurativa, laryngitis, mastitis, meningitis, myelitis myocarditis,myositis, nephritis, oophoritis, orchitis, osteitis, otitis,pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis,pleuritis, phlebitis, pneumonitis, pneumonia, proctitis, prostatitis,pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis,tendonitis, tonsillitis, uveitis, vaginitis, vasculitis, and vulvitis.

In yet other embodiments, the methods described herein can be used totreat a cancer, e.g., B-cell proliferative disorders, which include, butare not limited to diffuse large B cell lymphoma, follicular lymphoma,chronic lymphocytic lymphoma, chronic lymphocytic leukemia, B-cellprolymphocytic leukemia, lymphoplasmacytic lymphoma/Waldenstrammacroglobulinemia, splenic marginal zone lymphoma, plasma cell myeloma,plasmacytoma, extranodal marginal zone B cell lymphoma, nodal marginalzone B cell lymphoma, mantle cell lymphoma, mediastinal (thymic) large Bcell lymphoma, intravascular large B cell lymphoma, primary effusionlymphoma, burkitt lymphoma/leukemia, and lymphomatoid granulomatosis.

In further embodiments, the methods described herein can be used totreat thromboembolic disorders, which include, but are not limited tomyocardial infarct, angina pectoris (including unstable angina),reocclusions or restenoses after angioplasty or aortocoronary bypass,stroke, transitory ischemia, peripheral arterial occlusive disorders,pulmonary embolisms, and deep venous thromboses.

Hematological Malignancies

Disclosed herein, in certain embodiments, is a method for treating ahematological malignancy in an individual in need thereof, comprising:administering to the individual an amount of Compound 1.

In some embodiments, the hematological malignancy is a non-Hodgkin'slymphoma (NHL). In some embodiments, the hematological malignancy is achronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL),high risk CLL, or a non-CLL/SLL lymphoma. In some embodiments, thehematological malignancy is follicular lymphoma (FL), diffuse largeB-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), Waldenstrom'smacroglobulinemia, multiple myeloma (MM), marginal zone lymphoma,Burkitt's lymphoma, non-Burkitt high grade B cell lymphoma, orextranodal marginal zone B cell lymphoma. In some embodiments, thehematological malignancy is acute or chronic myelogenous (or myeloid)leukemia, myelodysplastic syndrome, acute lymphoblastic leukemia, orprecursor B-cell acute lymphoblastic leukemia. In some embodiments, thehematological malignancy is chronic lymphocytic leukemia (CLL). In someembodiments, the hematological malignancy is mantle cell lymphoma (MCL).In some embodiments, the hematological malignancy is diffuse largeB-cell lymphoma (DLBCL). In some embodiments, the hematologicalmalignancy is diffuse large B-cell lymphoma (DLBCL), ABC subtype. Insome embodiments, the hematological malignancy is diffuse large B-celllymphoma (DLBCL), GCB subtype. In some embodiments, the hematologicalmalignancy is Waldenstrom's macroglobulinemia (WM). In some embodiments,the hematological malignancy is multiple myeloma (MM). In someembodiments, the hematological malignancy is Burkitt's lymphoma. In someembodiments, the hematological malignancy is follicular lymphoma (FL).In some embodiments, the hematological malignancy is transformedfollicular lymphoma. In some embodiments, the hematological malignancyis marginal zone lymphoma.

In some embodiments, the hematological malignancy is relapsed orrefractory non-Hodgkin's lymphoma (NHL). In some embodiments, thehematological malignancy is relapsed or refractory diffuse large B-celllymphoma (DLBCL), relapsed or refractory mantle cell lymphoma (MCL),relapsed or refractory follicular lymphoma (FL), relapsed or refractoryCLL, relapsed or refractory SLL, relapsed or refractory multiplemyeloma, relapsed or refractory Waldenstrom's macroglobulinemia,relapsed or refractory multiple myeloma (MM), relapsed or refractorymarginal zone lymphoma, relapsed or refractory Burkitt's lymphoma,relapsed or refractory non-Burkitt high grade B cell lymphoma, relapsedor refractory extranodal marginal zone B cell lymphoma. In someembodiments, the hematological malignancy is a relapsed or refractoryacute or chronic myelogenous (or myeloid) leukemia, relapsed orrefractory myelodysplastic syndrome, relapsed or refractory acutelymphoblastic leukemia, or relapsed or refractory precursor B-cell acutelymphoblastic leukemia. In some embodiments, the hematologicalmalignancy is relapsed or refractory chronic lymphocytic leukemia (CLL).In some embodiments, the hematological malignancy is relapsed orrefractory mantle cell lymphoma (MCL). In some embodiments, thehematological malignancy is relapsed or refractory diffuse large B-celllymphoma (DLBCL). In some embodiments, the hematological malignancy isrelapsed or refractory diffuse large B-cell lymphoma (DLBCL), ABCsubtype. In some embodiments, the hematological malignancy is relapsedor refractory diffuse large B-cell lymphoma (DLBCL), GCB subtype. Insome embodiments, the hematological malignancy is relapsed or refractoryWaldenstrom's macroglobulinemia (WM). In some embodiments, thehematological malignancy is relapsed or refractory multiple myeloma(MM). In some embodiments, the hematological malignancy is relapsed orrefractory Burkitt's lymphoma. In some embodiments, the hematologicalmalignancy is relapsed or refractory follicular lymphoma (FL).

In some embodiments, the hematological malignancy is a hematologicalmalignancy that is classified as high-risk. In some embodiments, thehematological malignancy is high risk CLL or high risk SLL.

B-cell lymphoproliferative disorders (BCLDs) are neoplasms of the bloodand encompass, inter alia, non-Hodgkin lymphoma, multiple myeloma, andleukemia. BCLDs can originate either in the lymphatic tissues (as in thecase of lymphoma) or in the bone marrow (as in the case of leukemia andmyeloma), and they all are involved with the uncontrolled growth oflymphocytes or white blood cells. There are many subtypes of BCLD, e.g.,chronic lymphocytic leukemia (CLL) and non-Hodgkin lymphoma (NHL). Thedisease course and treatment of BCLD is dependent on the BCLD subtype;however, even within each subtype the clinical presentation, morphologicappearance, and response to therapy is heterogeneous.

Malignant lymphomas are neoplastic transformations of cells that residepredominantly within lymphoid tissues. Two groups of malignant lymphomasare Hodgkin's lymphoma and non-Hodgkin's lymphoma (NHL). Both types oflymphomas infiltrate reticuloendothelial tissues. However, they differin the neoplastic cell of origin, site of disease, presence of systemicsymptoms, and response to treatment (Freedman et al., “Non-Hodgkin'sLymphomas” Chapter 134, Cancer Medicine, (an approved publication of theAmerican Cancer Society, B.C. Decker Inc., Hamilton, Ontario, 2003).

Non-Hodgkin's Lymphomas

Disclosed herein, in certain embodiments, is a method for treating anon-Hodgkin's lymphoma in an individual in need thereof, comprising:administering to the individual an amount of Compound 1.

Further disclosed herein, in certain embodiments, is a method fortreating relapsed or refractory non-Hodgkin's lymphoma in an individualin need thereof, comprising: administering to the individual atherapeutically-effective amount of Compound 1. In some embodiments, thenon-Hodgkin's lymphoma is relapsed or refractory diffuse large B-celllymphoma (DLBCL), relapsed or refractory mantle cell lymphoma, relapsedor refractory follicular lymphoma, or relapsed or refractory CLL.

Non-Hodgkin lymphomas (NHL) are a diverse group of malignancies that arepredominately of B-cell origin. NHL may develop in any organs associatedwith lymphatic system such as spleen, lymph nodes or tonsils and canoccur at any age. NHL is often marked by enlarged lymph nodes, fever,and weight loss. NHL is classified as either B-cell or T-cell NHL.Lymphomas related to lymphoproliferative disorders following bone marrowor stem cell transplantation are usually B-cell NHL. In the WorkingFormulation classification scheme, NHL has been divided into low-,intermediate-, and high-grade categories by virtue of their naturalhistories (see “The Non-Hodgkin's Lymphoma Pathologic ClassificationProject,” Cancer 49(1982):2112-2135). The low-grade lymphomas areindolent, with a median survival of 5 to 10 years (Horning and Rosenberg(1984) N. Engl. J. Med. 311:1471-1475). Although chemotherapy can induceremissions in the majority of indolent lymphomas, cures are rare andmost patients eventually relapse, requiring further therapy. Theintermediate- and high-grade lymphomas are more aggressive tumors, butthey have a greater chance for cure with chemotherapy. However, asignificant proportion of these patients will relapse and requirefurther treatment.

A non-limiting list of the B-cell NHL includes Burkitt's lymphoma (e.g.,Endemic Burkitt's Lymphoma and Sporadic Burkitt's Lymphoma), CutaneousB-Cell Lymphoma, Cutaneous Marginal Zone Lymphoma (MZL), Diffuse LargeCell Lymphoma (DLBCL), Diffuse Mixed Small and Large Cell Lympoma,Diffuse Small Cleaved Cell, Diffuse Small Lymphocytic Lymphoma,Extranodal Marginal Zone B-cell lymphoma, follicular lymphoma,Follicular Small Cleaved Cell (Grade 1), Follicular Mixed Small Cleavedand Large Cell (Grade 2), Follicular Large Cell (Grade 3), IntravascularLarge B-Cell Lymphoma, Intravascular Lymphomatosis, Large CellImmunoblastic Lymphoma, Large Cell Lymphoma (LCL), LymphoblasticLymphoma, MALT Lymphoma, Mantle Cell Lymphoma (MCL), immunoblastic largecell lymphoma, precursor B-lymphoblastic lymphoma, mantle cell lymphoma,chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL),extranodal marginal zone B-cell lymphoma-mucosa-associated lymphoidtissue (MALT) lymphoma, Mediastinal Large B-Cell Lymphoma, nodalmarginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma,primary mediastinal B-cell lymphoma, lymphoplasmocytic lymphoma, hairycell leukemia, Waldenstrom's Macroglobulinemia, and primary centralnervous system (CNS) lymphoma. Additional non-Hodgkin's lymphomas arecontemplated within the scope of the present invention and apparent tothose of ordinary skill in the art.

DLBCL

Disclosed herein, in certain embodiments, is a method for treating aDLCBL in an individual in need thereof, comprising: administering to theindividual an amount of Compound 1. Further disclosed herein, in certainembodiments, is a method for treating relapsed or refractory DLCBL in anindividual in need thereof, comprising: administering to the individuala therapeutically-effective amount of Compound 1.

As used herein, the term “Diffuse large B-cell lymphoma (DLBCL)” refersto a neoplasm of the germinal center B lymphocytes with a diffuse growthpattern and a high-intermediate proliferation index. DLBCLs representapproximately 30% of all lymphomas and may present with severalmorphological variants including the centroblastic, immunoblastic,T-cell/histiocyte rich, anaplastic and plasmoblastic subtypes. Genetictests have shown that there are different subtypes of DLBCL. Thesesubtypes seem to have different outlooks (prognoses) and responses totreatment. DLBCL can affect any age group but occurs mostly in olderpeople (the average age is mid-60s).

Disclosed herein, in certain embodiments, is a method for treatingdiffuse large B-cell lymphoma, activated B cell-like subtype(ABC-DLBCL), in an individual in need thereof, comprising: administeringto the individual an irreversible Btk inhibitor in an amount from 300mg/day up to, and including, 1000 mg/day. The ABC subtype of diffuselarge B-cell lymphoma (ABC-DLBCL) is thought to arise from post germinalcenter B cells that are arrested during plasmatic differentiation. TheABC subtype of DLBCL (ABC-DLBCL) accounts for approximately 30% totalDLBCL diagnoses. It is considered the least curable of the DLBCLmolecular subtypes and, as such, patients diagnosed with the ABC-DLBCLtypically display significantly reduced survival rates compared withindividuals with other types of DLCBL. ABC-DLBCL is most commonlyassociated with chromosomal translocations deregulating the germinalcenter master regulator BCL6 and with mutations inactivating the PRDM1gene, which encodes a transcriptional repressor required for plasma celldifferentiation.

A particularly relevant signaling pathway in the pathogenesis ofABC-DLBCL is the one mediated by the nuclear factor (NF)-κBtranscription complex. The NF-κB family comprises 5 members (p50, p52,p65, c-rel and RelB) that form homo- and heterodimers and function astranscriptional factors to mediate a variety of proliferation,apoptosis, inflammatory and immune responses and are critical for normalB-cell development and survival. NF-κB is widely used by eukaryoticcells as a regulator of genes that control cell proliferation and cellsurvival. As such, many different types of human tumors havemisregulated NF-κB: that is, NF-κB is constitutively active. ActiveNF-κB turns on the expression of genes that keep the cell proliferatingand protect the cell from conditions that would otherwise cause it todie via apoptosis.

The dependence of ABC DLBCLs on NF-kB depends on a signaling pathwayupstream of IkB kinase comprised of CARD11, BCL10 and MALT1 (the CBMcomplex). Interference with the CBM pathway extinguishes NF-kB signalingin ABC DLBCL cells and induces apoptosis. The molecular basis forconstitutive activity of the NF-kB pathway is a subject of currentinvestigation but some somatic alterations to the genome of ABC DLBCLsclearly invoke this pathway. For example, somatic mutations of thecoiled-coil domain of CARD11 in DLBCL render this signaling scaffoldprotein able to spontaneously nucleate protein-protein interaction withMALT1 and BCL10, causing IKK activity and NF-kB activation. Constitutiveactivity of the B cell receptor signaling pathway has been implicated inthe activation of NF-kB in ABC DLBCLs with wild type CARD11, and this isassociated with mutations within the cytoplasmic tails of the B cellreceptor subunits CD79A and CD79B. Oncogenic activating mutations in thesignaling adapter MYD88 activate NF-kB and synergize with B cellreceptor signaling in sustaining the survival of ABC DLBCL cells. Inaddition, inactivating mutations in a negative regulator of the NF-kBpathway, A20, occur almost exclusively in ABC DLBCL.

Indeed, genetic alterations affecting multiple components of the NF-κBsignaling pathway have been recently identified in more than 50% ofABC-DLBCL patients, where these lesions promote constitutive NF-κBactivation, thereby contributing to lymphoma growth. These includemutations of CARD11 (˜10% of the cases), a lymphocyte-specificcytoplasmic scaffolding protein that-together with MALT1 and BCL10-formsthe BCR signalosome, which relays signals from antigen receptors to thedownstream mediators of NF-κB activation. An even larger fraction ofcases (˜30%) carry biallelic genetic lesions inactivating the negativeNF-κB regulator A20. Further, high levels of expression of NF-κB targetgenes have been observed in ABC-DLBCL tumor samples. See, e.g., U. Kleinet al., (2008), Nature Reviews Immunology 8:22-23; R. E. Davis et al.,(2001), Journal of Experimental Medicine 194:1861-1874; G. Lentz et al.,(2008), Science 319:1676-1679; M. Compagno et al., (2009), Nature459:712-721; and L. Srinivasan et al., (2009), Cell 139:573-586).

DLBCL cells of the ABC subtype, such as OCI-Ly10, have chronic activeBCR signalling and are very sensitive to the Btk inhibitor describedherein. The irreversible Btk inhibitor described herein potently andirreversibly inhibits the growth of OCI-Ly10 (EC₅₀ continuousexposure=10 nM, EC₅₀ 1 hour pulse=50 nM). In addition, induction ofapoptosis, as shown by capsase activation, Annexin-V flow cytometry andincrease in sub-GO fraction is observed in OCILy10. Both sensitive andresistant cells express Btk at similar levels, and the active site ofBtk is fully occupied by the inhibitor in both as shown using afluorescently labeled affinity probe. OCI-Ly10 cells are shown to havechronically active BCR signalling to NF-kB which is dose dependentlyinhibited by the Btk inhibitors described herein. The activity of Btkinhibitors in the cell lines studied herein are also characterized bycomparing signal transduction profiles (Btk, PLCγ, ERK, NF-κB, AKT),cytokine secretion profiles and mRNA expression profiles, both with andwithout BCR stimulation, and observed significant differences in theseprofiles that lead to clinical biomarkers that identify the mostsensitive patient populations to Btk inhibitor treatment. See U.S. Pat.No. 7,711,492 and Staudt et al., Nature, Vol. 463, Jan. 7, 2010, pp.88-92, the contents of which are incorporated by reference in theirentirety.

Follicular Lymphoma

Disclosed herein, in certain embodiments, is a method for treating afollicular lymphoma in an individual in need thereof, comprising:administering to the individual an amount of Compound 1. Furtherdisclosed herein, in certain embodiments, is a method for treatingrelapsed or refractory follicular lymphoma in an individual in needthereof, comprising: administering to the individual atherapeutically-effective amount of Compound 1.

As used herein, the term “follicular lymphoma” refers to any of severaltypes of non-Hodgkin's lymphoma in which the lymphomatous cells areclustered into nodules or follicles. The term follicular is used becausethe cells tend to grow in a circular, or nodular, pattern in lymphnodes. The average age for people with this lymphoma is about 60.

CLL/SLL

Disclosed herein, in certain embodiments, is a method for treating a CLLor SLL in an individual in need thereof, comprising: administering tothe individual an amount of Compound 1. Further disclosed herein, incertain embodiments, is a method for treating relapsed or refractory CLLor SLL in an individual in need thereof, comprising: administering tothe individual a therapeutically-effective amount of Compound 1.

Chronic lymphocytic leukemia and small lymphocytic lymphoma (CLL/SLL)are commonly thought as the same disease with slightly differentmanifestations. Where the cancerous cells gather determines whether itis called CLL or SLL. When the cancer cells are primarily found in thelymph nodes, lima bean shaped structures of the lymphatic system (asystem primarily of tiny vessels found in the body), it is called SLL.SLL accounts for about 5% to 10% of all lymphomas. When most of thecancer cells are in the bloodstream and the bone marrow, it is calledCLL.

Both CLL and SLL are slow-growing diseases, although CLL, which is muchmore common, tends to grow slower. CLL and SLL are treated the same way.They are usually not considered curable with standard treatments, butdepending on the stage and growth rate of the disease, most patientslive longer than 10 years. Occasionally over time, these slow-growinglymphomas may transform into a more aggressive type of lymphoma.

Chronic lymphoid leukemia (CLL) is the most common type of leukemia. Itis estimated that 100,760 people in the United States are living with orare in remission from CLL. Most (>75%) people newly diagnosed with CLLare over the age of 50. Currently CLL treatment focuses on controllingthe disease and its symptoms rather than on an outright cure. CLL istreated by chemotherapy, radiation therapy, biological therapy, or bonemarrow transplantation. Symptoms are sometimes treated surgically(splenectomy removal of enlarged spleen) or by radiation therapy(“de-bulking” swollen lymph nodes). Though CLL progresses slowly in mostcases, it is considered generally incurable. Certain CLLs are classifiedas high-risk. As used herein, “high risk CLL” means CLL characterized byat least one of the following 1) 17p13-; 2) 11q22-; 3) unmutated IgVHtogether with ZAP-70+ and/or CD38+; or 4) trisomy 12.

CLL treatment is typically administered when the patient's clinicalsymptoms or blood counts indicate that the disease has progressed to apoint where it may affect the patient's quality of life.

Small lymphocytic leukemia (SLL) is very similar to CLL described supra,and is also a cancer of B-cells. In SLL the abnormal lymphocytes mainlyaffect the lymph nodes. However, in CLL the abnormal cells mainly affectthe blood and the bone marrow. The spleen may be affected in bothconditions. SLL accounts for about 1 in 25 of all cases of non-Hodgkinlymphoma. It can occur at any time from young adulthood to old age, butis rare under the age of 50. SLL is considered an indolent lymphoma.This means that the disease progresses very slowly, and patients tend tolive many years after diagnosis. However, most patients are diagnosedwith advanced disease, and although SLL responds well to a variety ofchemotherapy drugs, it is generally considered to be incurable. Althoughsome cancers tend to occur more often in one gender or the other, casesand deaths due to SLL are evenly split between men and women. Theaverage age at the time of diagnosis is 60 years.

Although SLL is indolent, it is persistently progressive. The usualpattern of this disease is one of high response rates to radiationtherapy and/or chemotherapy, with a period of disease remission. This isfollowed months or years later by an inevitable relapse. Re-treatmentleads to a response again, but again the disease will relapse. Thismeans that although the short-term prognosis of SLL is quite good, overtime, many patients develop fatal complications of recurrent disease.Considering the age of the individuals typically diagnosed with CLL andSLL, there is a need in the art for a simple and effective treatment ofthe disease with minimum side-effects that do not impede on thepatient's quality of life. The instant invention fulfills this longstanding need in the art.

Mantle Cell Lymphoma

Disclosed herein, in certain embodiments, is a method for treating aMantle cell lymphoma in an individual in need thereof, comprising:administering to the individual an amount of Compound 1. Furtherdisclosed herein, in certain embodiments, is a method for treatingrelapsed or refractory Mantle cell lymphoma in an individual in needthereof, comprising: administering to the individual atherapeutically-effective amount of Compound 1.

As used herein, the term, “Mantle cell lymphoma” refers to a subtype ofB-cell lymphoma, due to CD5 positive antigen-naive pregerminal centerB-cell within the mantle zone that surrounds normal germinal centerfollicles. MCL cells generally over-express cyclin D1 due to a t(11:14)chromosomal translocation in the DNA. More specifically, thetranslocation is at t(11;14)(q13;q32). Only about 5% of lymphomas are ofthis type. The cells are small to medium in size. Men are affected mostoften. The average age of patients is in the early 60s. The lymphoma isusually widespread when it is diagnosed, involving lymph nodes, bonemarrow, and, very often, the spleen. Mantle cell lymphoma is not a veryfast growing lymphoma, but is difficult to treat.

Marginal Zone B-Cell Lymphoma

Disclosed herein, in certain embodiments, is a method for treating amarginal zone B-cell lymphoma in an individual in need thereof,comprising: administering to the individual an amount of Compound 1.Further disclosed herein, in certain embodiments, is a method fortreating relapsed or refractory marginal zone B-cell lymphoma in anindividual in need thereof, comprising: administering to the individuala therapeutically-effective amount of Compound 1.

As used herein, the term “marginal zone B-cell lymphoma” refers to agroup of related B-cell neoplasms that involve the lymphoid tissues inthe marginal zone, the patchy area outside the follicular mantle zone.Marginal zone lymphomas account for about 5% to 10% of lymphomas. Thecells in these lymphomas look small under the microscope. There are 3main types of marginal zone lymphomas including extranodal marginal zoneB-cell lymphomas, nodal marginal zone B-cell lymphoma, and splenicmarginal zone lymphoma.

MALT

Disclosed herein, in certain embodiments, is a method for treating aMALT in an individual in need thereof, comprising: administering to theindividual an amount of Compound 1. Further disclosed herein, in certainembodiments, is a method for treating relapsed or refractory MALT in anindividual in need thereof, comprising: administering to the individuala therapeutically-effective amount of Compound 1.

The term “mucosa-associated lymphoid tissue (MALT) lymphoma”, as usedherein, refers to extranodal manifestations of marginal-zone lymphomas.Most MALT lymphoma are a low grade, although a minority either manifestinitially as intermediate-grade non-Hodgkin lymphoma (NHL) or evolvefrom the low-grade form. Most of the MALT lymphoma occur in the stomach,and roughly 70% of gastric MALT lymphoma are associated withHelicobacter pylori infection. Several cytogenetic abnormalities havebeen identified, the most common being trisomy 3 or t(11;18). Many ofthese other MALT lymphoma have also been linked to infections withbacteria or viruses. The average age of patients with MALT lymphoma isabout 60.

Nodal Marginal Zone B-Cell Lymphoma

Disclosed herein, in certain embodiments, is a method for treating anodal marginal zone B-cell lymphoma in an individual in need thereof,comprising: administering to the individual an amount of Compound 1.Further disclosed herein, in certain embodiments, is a method fortreating relapsed or refractory nodal marginal zone B-cell lymphoma inan individual in need thereof, comprising: administering to theindividual a therapeutically-effective amount of Compound 1.

The term “nodal marginal zone B-cell lymphoma” refers to an indolentB-cell lymphoma that is found mostly in the lymph nodes. The disease israre and only accounts for 1% of all Non-Hodgkin's Lymphomas (NHL). Itis most commonly diagnosed in older patients, with women moresusceptible than men. The disease is classified as a marginal zonelymphoma because the mutation occurs in the marginal zone of theB-cells. Due to its confinement in the lymph nodes, this disease is alsoclassified as nodal.

Splenic Marginal Zone B-Cell Lymphoma

Disclosed herein, in certain embodiments, is a method for treating asplenic marginal zone B-cell lymphoma in an individual in need thereof,comprising: administering to the individual an amount of Compound 1.Further disclosed herein, in certain embodiments, is a method fortreating relapsed or refractory splenic marginal zone B-cell lymphoma inan individual in need thereof, comprising: administering to theindividual a therapeutically-effective amount of Compound 1.

The term “splenic marginal zone B-cell lymphoma” refers to specificlow-grade small B-cell lymphoma that is incorporated in the World HealthOrganization classification. Characteristic features are splenomegaly,moderate lymphocytosis with villous morphology, intrasinusoidal patternof involvement of various organs, especially bone marrow, and relativeindolent course. Tumor progression with increase of blastic forms andaggressive behavior are observed in a minority of patients. Molecularand cytogenetic studies have shown heterogeneous results probablybecause of the lack of standardized diagnostic criteria.

Burkitt Lymphoma

Disclosed herein, in certain embodiments, is a method for treating aBurkitt lymphoma in an individual in need thereof, comprising:administering to the individual an amount of Compound 1. Furtherdisclosed herein, in certain embodiments, is a method for treatingrelapsed or refractory Burkitt lymphoma in an individual in needthereof, comprising: administering to the individual atherapeutically-effective amount of Compound 1.

The term “Burkitt lymphoma” refers to a type of Non-Hodgkin Lymphoma(NHL) that commonly affects children. It is a highly aggressive type ofB-cell lymphoma that often starts and involves body parts other thanlymph nodes. In spite of its fast-growing nature, Burkitt's lymphoma isoften curable with modern intensive therapies. There are two broad typesof Burkitt's lymphoma—the sporadic and the endemic varieties:

Endemic Burkitt's lymphoma: The disease involves children much more thanadults, and is related to Epstein Barr Virus (EBV) infection in 95%cases. It occurs primarily is equatorial Africa, where about half of allchildhood cancers are Burkitt's lymphoma. It characteristically has ahigh chance of involving the jawbone, a rather distinctive feature thatis rare in sporadic Burkitt's. It also commonly involves the abdomen.

Sporadic Burkitt's lymphoma: The type of Burkitt's lymphoma that affectsthe rest of the world, including Europe and the Americas is the sporadictype. Here too, it's mainly a disease in children. The link betweenEpstein Barr Virus (EBV) is not as strong as with the endemic variety,though direct evidence of EBV infection is present in one out of fivepatients. More than the involvement of lymph nodes, it is the abdomenthat is notably affected in more than 90% of the children. Bone marrowinvolvement is more common than in the sporadic variety.

Waldenstrom Macroglobulinemia

Disclosed herein, in certain embodiments, is a method for treating aWaldenstrom macroglobulinemia in an individual in need thereof,comprising: administering to the individual an amount of Compound 1.Further disclosed herein, in certain embodiments, is a method fortreating relapsed or refractory Waldenstrom macroglobulinemia in anindividual in need thereof, comprising: administering to the individuala therapeutically-effective amount of Compound 1.

The term “Waldenstrom macroglobulinemia”, also known aslymphoplasmacytic lymphoma, is cancer involving a subtype of white bloodcells called lymphocytes. It is characterized by an uncontrolled clonalproliferation of terminally differentiated B lymphocytes. It is alsocharacterized by the lymphoma cells making an antibody calledimmunoglobulin M (IgM). The IgM antibodies circulate in the blood inlarge amounts, and cause the liquid part of the blood to thicken, likesyrup. This can lead to decreased blood flow to many organs, which cancause problems with vision (because of poor circulation in blood vesselsin the back of the eyes) and neurological problems (such as headache,dizziness, and confusion) caused by poor blood flow within the brain.Other symptoms can include feeling tired and weak, and a tendency tobleed easily. The underlying etiology is not fully understood but anumber of risk factors have been identified, including the locus 6p21.3on chromosome 6. There is a 2- to 3-fold risk increase of developing WMin people with a personal history of autoimmune diseases withautoantibodies and particularly elevated risks associated withhepatitis, human immunodeficiency virus, and rickettsiosis.

Multiple Myeloma

Disclosed herein, in certain embodiments, is a method for treating amyeloma in an individual in need thereof, comprising: administering tothe individual an amount of Compound 1. Further disclosed herein, incertain embodiments, is a method for treating relapsed or refractorymyeloma in an individual in need thereof, comprising: administering tothe individual a therapeutically-effective amount of Compound 1.

Multiple myeloma, also known as MM, myeloma, plasma cell myeloma, or asKahler's disease (after Otto Kahler) is a cancer of the white bloodcells known as plasma cells. A type of B cell, plasma cells are acrucial part of the immune system responsible for the production ofantibodies in humans and other vertebrates. They are produced in thebone marrow and are transported through the lymphatic system.

Leukemia

Disclosed herein, in certain embodiments, is a method for treating aleukemia in an individual in need thereof, comprising: administering tothe individual an amount of Compound 1. Further disclosed herein, incertain embodiments, is a method for treating relapsed or refractoryleukemia in an individual in need thereof, comprising: administering tothe individual a therapeutically-effective amount of Compound 1.

Leukemia is a cancer of the blood or bone marrow characterized by anabnormal increase of blood cells, usually leukocytes (white bloodcells). Leukemia is a broad term covering a spectrum of diseases. Thefirst division is between its acute and chronic forms: (i) acuteleukemia is characterized by the rapid increase of immature blood cells.This crowding makes the bone marrow unable to produce healthy bloodcells. Immediate treatment is required in acute leukemia due to therapid progression and accumulation of the malignant cells, which thenspill over into the bloodstream and spread to other organs of the body.Acute forms of leukemia are the most common forms of leukemia inchildren; (ii) chronic leukemia is distinguished by the excessive buildup of relatively mature, but still abnormal, white blood cells.Typically taking months or years to progress, the cells are produced ata much higher rate than normal cells, resulting in many abnormal whiteblood cells in the blood. Chronic leukemia mostly occurs in olderpeople, but can theoretically occur in any age group. Additionally, thediseases are subdivided according to which kind of blood cell isaffected. This split divides leukemias into lymphoblastic or lymphocyticleukemias and myeloid or myelogenous leukemias: (i) lymphoblastic orlymphocytic leukemias, the cancerous change takes place in a type ofmarrow cell that normally goes on to form lymphocytes, which areinfection-fighting immune system cells; (ii) myeloid or myelogenousleukemias, the cancerous change takes place in a type of marrow cellthat normally goes on to form red blood cells, some other types of whitecells, and platelets.

Within these main categories, there are several subcategories including,but not limited to, Acute lymphoblastic leukemia (ALL), precursor B-cellacute lymphoblastic leukemia (precursor B-ALL; also called precursorB-lymphoblastic leukemia), Acute myelogenous leukemia (AML), Chronicmyelogenous leukemia (CML), and Hairy cell leukemia (HCL). Accordingly,disclosed herein, in certain embodiments, is a method for treating Acutelymphoblastic leukemia (ALL), precursor B-cell acute lymphoblasticleukemia (precursor B-ALL; also called precursor B-lymphoblasticleukemia), Acute myelogenous leukemia (AML), Chronic myelogenousleukemia (CML), or Hairy cell leukemia (HCL) in an individual in needthereof, comprising: administering to the individual an amount ofCompound 1. In some embodiments, the leukemia is a relapsed orrefractory leukemia. In some embodiments, the leukemia is a relapsed orrefractory Acute lymphoblastic leukemia (ALL), relapsed or refractoryprecursor B-cell acute lymphoblastic leukemia (precursor B-ALL; alsocalled precursor B-lymphoblastic leukemia), relapsed or refractory Acutemyelogenous leukemia (AML), relapsed or refractory Chronic myelogenousleukemia (CML), or relapsed or refractory Hairy cell leukemia (HCL).

Symptoms, diagnostic tests, and prognostic tests for each of theabove-mentioned conditions are known. See, e.g., Harrison's Principlesof Internal Medicine©,” 16th ed., 2004, The McGraw-Hill Companies, Inc.Dey et al. (2006), Cytojournal 3(24), and the “Revised European AmericanLymphoma” (REAL) classification system (see, e.g., the websitemaintained by the National Cancer Institute).

A number of animal models of are useful for establishing a range oftherapeutically effective doses of irreversible Btk inhibitor compounds,such as Compound 1, for treating any of the foregoing diseases.

The therapeutic efficacy of Compound 1 for any one of the foregoingdiseases can be optimized during a course of treatment. For example, asubject being treated can undergo a diagnostic evaluation to correlatethe relief of disease symptoms or pathologies to inhibition of in vivoBtk activity achieved by administering a given dose of Compound 1.Cellular assays known in the art can be used to determine in vivoactivity of Btk in the presence or absence of an irreversible Btkinhibitor. For example, since activated Btk is phosphorylated attyrosine 223 (Y223) and tyrosine 551 (Y551), phospho-specificimmunocytochemical staining of P-Y223 or P-Y551-positive cells can beused to detect or quantify activation of Bkt in a population of cells(e.g., by FACS analysis of stained vs unstained cells). See, e.g.,Nisitani et al. (1999), Proc. Nat. Acad. Sci, USA 96:2221-2226. Thus,the amount of the Btk inhibitor compound that is administered to asubject can be increased or decreased as needed so as to maintain alevel of Btk inhibition optimal for treating the subject's diseasestate.

Compound 1 can irreversibly inhibit Btk and may be used to treat mammalssuffering from Bruton's tyrosine kinase-dependent or Bruton's tyrosinekinase mediated conditions or diseases, including, but not limited to,cancer, autoimmune and other inflammatory diseases. Compound 1 has shownefficacy is a wide variety of diseases and conditions that are describedherein.

In some embodiments, Compound 1 is used for the manufacture of amedicament for treating any of the foregoing conditions (e.g.,autoimmune diseases, inflammatory diseases, allergy disorders, B-cellproliferative disorders, or thromboembolic disorders).

Compound 1, and Pharmaceutically Acceptable Salts Thereof

The Btk inhibitor compound described herein (i.e. Compound 1) isselective for Btk and kinases having a cysteine residue in an amino acidsequence position of the tyrosine kinase that is homologous to the aminoacid sequence position of cysteine 481 in Btk. The Btk inhibitorcompound can form a covalent bond with Cys 481 of Btk (e.g., via aMichael reaction).

“Compound 1” or“1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one”or“1-{(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl}prop-2-en-1-one”or “2-Propen-1-one,1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl-”or ibrutinib or any other suitable name refers to the compound with thefollowing structure:

A wide variety of pharmaceutically acceptable salts is formed fromCompound 1 and includes:

-   -   acid addition salts formed by reacting Compound 1 with an        organic acid, which includes aliphatic mono- and dicarboxylic        acids, phenyl-substituted alkanoic acids, hydroxyl alkanoic        acids, alkanedioic acids, aromatic acids, aliphatic and aromatic        sulfonic acids, amino acids, etc. and include, for example,        acetic acid, trifluoroacetic acid, propionic acid, glycolic        acid, pyruvic acid, oxalic acid, maleic acid, malonic acid,        succinic acid, fumaric acid, tartaric acid, critic acid, benzoic        acid, cinnamic acid, mandelic acid, methanesulfonic acid,        ethanesulfonica acid, p-toluenesulfonic acid, salicylic acid,        and the like;    -   acid addition salts formed by reacting Compound 1 with an        inorganic acid, which includes hydrochloric acid, hydrobromic        acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic        acid, hydrofluoric acid, phosphorous acid, and the like.

The term “pharmaceutically acceptable salts” in reference to Compound 1refers to a salt of Compound 1, which does not cause significantirritation to a mammal to which it is administered and does notsubstantially abrogate the biological activity and properties of thecompound.

It should be understood that a reference to a pharmaceuticallyacceptable salt includes the solvent addition forms (solvates). Solvatescontain either stoichiometric or non-stoichiometric amounts of asolvent, and are formed during the process of product formation orisolation with pharmaceutically acceptable solvents such as water,ethanol, methanol, methyl tert-butyl ether (MTBE), diisopropyl ether(DIPE), ethyl acetate, isopropyl acetate, isopropyl alcohol, methylisobutyl ketone (MIBK), methyl ethyl ketone (MEK), acetone,nitromethane, tetrahydrofuran (THF), dichloromethane (DCM), dioxane,heptanes, toluene, anisole, acetonitrile, and the like. In one aspect,solvates are formed using, but not limited to, Class 3 solvent(s).Categories of solvents are defined in, for example, the InternationalConference on Harmonization of Technical Requirements for Registrationof Pharmaceuticals for Human Use (ICH), “Impurities: Guidelines forResidual Solvents, Q3C(R3), (November 2005). Hydrates are formed whenthe solvent is water, or alcoholates are formed when the solvent isalcohol. In some embodiments, solvates of Compound 1, orpharmaceutically acceptable salts thereof, are conveniently prepared orformed during the processes described herein. In some embodiments,solvates of Compound 1 are anhydrous. In some embodiments, Compound 1,or pharmaceutically acceptable salts thereof, exist in unsolvated form.In some embodiments, Compound 1, or pharmaceutically acceptable saltsthereof, exist in unsolvated form and are anhydrous.

In yet other embodiments, Compound 1, or a pharmaceutically acceptablesalt thereof, is prepared in various forms, including but not limitedto, amorphous phase, crystalline forms, milled forms andnano-particulate forms. In some embodiments, Compound 1, or apharmaceutically acceptable salt thereof, is amorphous. In someembodiments, Compound 1, or a pharmaceutically acceptable salt thereof,is amorphous and anhydrous. In some embodiments, Compound 1, or apharmaceutically acceptable salt thereof, is crystalline. In someembodiments, Compound 1, or a pharmaceutically acceptable salt thereof,is crystalline and anhydrous.

In some embodiments, Compound 1 is prepared as outlined in U.S. Pat. No.7,514,444.

Amorphous Compound 1

In some embodiments, Compound 1 is amorphous and anhydrous. In someembodiments, Compound 1 is amorphous. In some embodiments, amorphousCompound 1 has an X-Ray Powder Diffraction (XRPD) pattern showing a lackof crystallinity.

Compound 1, Form A

In some embodiments, Compound 1 is crystalline. In some embodiments,Compound 1 is crystalline Form A. Crystalline Form A of Compound 1 ischaracterized as having at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 1;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 5.7±0.1° 2-Theta, 13.6±0.1° 2-Theta,        16.1±0.1° 2-Theta, 18.9±0.1° 2-Theta, 21.3±0.1° 2-Theta, and        21.6±0.1° 2-Theta;    -   (c) substantially the same X-ray powder diffraction (XRPD)        pattern post storage at 40° C. and 75% RH for at least a week;    -   (d) substantially the same X-ray powder diffraction (XRPD)        pattern post storage at 25° C. and 97% RH for at least a week;    -   (e) Infrared (IR) spectrum substantially similar to the one set        forth in FIG. 2;    -   (f) Infrared (IR) spectrum weak peaks at about 1584 cm⁻¹, about        1240 cm⁻¹, about 1147 cm⁻¹, about 1134 cm⁻¹, about 1099 cm⁻¹,        and about 1067 cm⁻¹;    -   (g) a DSC thermogram substantially similar to the one set forth        in FIG. 3;    -   (h) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 4;    -   (i) a DSC thermogram with an endotherm having an onset at about        154° C. and a peak at about 157° C. and an exotherm at about        159° C.;    -   (j) non-hygroscopicity;    -   (k) an observed aqueous solubility of about 0.013 mg/mL at about        pH 8;    -   or    -   (l) combinations thereof.

In some embodiments, Form A of Compound 1 is characterized as having atleast two of the properties selected from (a) to (k). In someembodiments, Form A of Compound 1 is characterized as having at leastthree of the properties selected from (a) to (k). In some embodiments,Form A of Compound 1 is characterized as having at least four of theproperties selected from (a) to (k). In some embodiments, Form A ofCompound 1 is characterized as having at least five of the propertiesselected from (a) to (k). In some embodiments, Form A of Compound 1 ischaracterized as having at least six of the properties selected from (a)to (k). In some embodiments, Form A of Compound 1 is characterized ashaving at least seven of the properties selected from (a) to (k). Insome embodiments, Form A of Compound 1 is characterized as having atleast eight of the properties selected from (a) to (k). In someembodiments, Form A of Compound 1 is characterized as having at leastnine of the properties selected from (a) to (k). In some embodiments,Form A of Compound 1 is characterized as having at least ten of theproperties selected from (a) to (k). In some embodiments, Form A ofCompound 1 is characterized as having properties (a) to (k).

In some embodiments, Form A has an X-Ray powder diffraction (XRPD)pattern substantially the same as shown in FIG. 1. In some embodiments,Form A has an X-ray powder diffraction (XRPD) pattern withcharacteristic peaks at 5.7±0.1° 2-Theta, 13.6±0.1° 2-Theta, 16.1±0.1°2-Theta, 18.9±0.1° 2-Theta, 21.3±0.1° 2-Theta, and 21.6±0.1° 2-Theta. Insome embodiments, Form A has substantially the same X-ray powderdiffraction (XRPD) pattern post storage at 40° C. and 75% RH for atleast a week. In some embodiments, Form A has substantially the sameX-ray powder diffraction (XRPD) pattern post storage at 25° C. and 97%RH for at least a week.

In some embodiments, Form A has an Infrared (IR) spectrum substantiallysimilar to the one set forth in FIG. 2. In some embodiments, Form A hasan Infrared (IR) spectrum weak peaks at about 1584 cm⁻¹, about 1240cm⁻¹, about 1147 cm⁻¹, about 1134 cm⁻¹, about 1099 cm⁻¹, and about 1067cm⁻¹.

In some embodiments, Form A has a DSC thermogram substantially similarto the one set forth in FIG. 3. In some embodiments, Form A has athermo-gravimetric analysis (TGA) thermogram substantially similar tothe one set forth in FIG. 4. In some embodiments, Form A has a DSCthermogram with an endotherm having an onset at about 154° C. and a peakat about 157° C. and an exotherm at about 159° C.

In some embodiments, Form A has non-hygroscopicity.

In some embodiments, Form A has an observed aqueous solubility of about0.013 mg/mL at about pH 8.

In some embodiments, Form A was obtained from ethyl acetate, isopropylacetate, tetrahydrofuran, methyl isobutyl ketone (MIBK), methyl ethylketone (MEK), nitromethane, methanol, ethanol, acetonitrile, dioxane,methyl tert-butyl ether (MTBE), anisole, acetone, heptanes, amethanol/water mixture or an acetone/heptane mixture. In someembodiments, Form A was obtained from ethyl acetate, isopropyl acetate,tetrahydrofuran, methyl isobutyl ketone (MIBK), methyl ethyl ketone(MEK), nitromethane, methanol, ethanol, acetonitrile, dioxane, methyltert-butyl ether (MTBE), anisole, acetone, heptanes, or anacetone/heptane mixture.

In some embodiments, Form A is unsolvated. In some embodiments, Form Ais anhydrous.

Compound 1, Form B

In some embodiments, Compound 1 is crystalline. In some embodiments,Compound 1 is crystalline Form B. Crystalline Form B of Compound 1 ischaracterized as having at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 5;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 5.2±0.1° 2-Theta, 10.2±0.1° 2-Theta,        16.5±0.1° 2-Theta, 18.5±0.1° 2-Theta, and 20.8±0.1° 2-Theta;    -   (c) substantially the same X-ray powder diffraction (XRPD)        pattern post storage at 40° C. and 75% RH for at least a week;    -   (d) substantially the same X-ray powder diffraction (XRPD)        pattern post storage at 25° C. and 97% RH for at least a week;    -   (e) Infrared (IR) spectrum substantially similar to the one set        forth in FIG. 6;    -   (f) Infrared (IR) spectrum weak peaks at about 1586 cm⁻¹, about        1573 cm⁻¹, about 1562 cm⁻¹, about 1229 cm⁻¹, about 1141 cm⁻¹,        about 1103 cm⁻¹, about 1056 cm⁻¹, and about 1033 cm⁻¹;    -   (g) a DSC thermogram substantially similar to the one set forth        in FIG. 7;    -   (h) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 8;    -   (i) a DSC thermogram with an endotherm having an onset at about        99-106° C. and a peak at about 115-118° C.;    -   (j) an observed aqueous solubility of about 0.0096 mg/mL at a pH        of about 7.42;    -   or    -   (k) combinations thereof.

In some embodiments, Form B of Compound 1 is characterized as having atleast two of the properties selected from (a) to (j). In someembodiments, Form B of Compound 1 is characterized as having at leastthree of the properties selected from (a) to (j). In some embodiments,Form B of Compound 1 is characterized as having at least four of theproperties selected from (a) to (j). In some embodiments, Form B ofCompound 1 is characterized as having at least five of the propertiesselected from (a) to (j). In some embodiments, Form B of Compound 1 ischaracterized as having at least six of the properties selected from (a)to (j). In some embodiments, Form B of Compound 1 is characterized ashaving at least seven of the properties selected from (a) to (j). Insome embodiments, Form B of Compound 1 is characterized as having atleast eight of the properties selected from (a) to (j). In someembodiments, Form B of Compound 1 is characterized as having at leastnine of the properties selected from (a) to (j). In some embodiments,Form B of Compound 1 is characterized as having properties (a) to (j).

In some embodiments, Form B has an X-Ray powder diffraction (XRPD)pattern substantially the same as shown in FIG. 5. In some embodiments,Form B has an X-ray powder diffraction (XRPD) pattern withcharacteristic peaks at 5.2±0.1° 2-Theta, 10.2±0.1° 2-Theta, 16.5±0.1°2-Theta, 18.5±0.1° 2-Theta, and 20.8±0.1° 2-Theta. In some embodiments,Form B has substantially the same X-ray powder diffraction (XRPD)pattern post storage at 40° C. and 75% RH for at least a week. In someembodiments, Form B has substantially the same X-ray powder diffraction(XRPD) pattern post storage at 25° C. and 97% RH for at least a week.

In some embodiments, Form B has an Infrared (IR) spectrum substantiallysimilar to the one set forth in FIG. 6. In some embodiments, Form B hasan Infrared (IR) spectrum weak peaks at about 1586 cm⁻¹, about 1573cm⁻¹, about 1562 cm⁻¹, about 1229 cm⁻¹, about 1141 cm¹, about 1103 cm⁻¹,about 1056 cm⁻¹, and about 1033 cm⁻¹.

In some embodiments, Form B has a DSC thermogram substantially similarto the one set forth in FIG. 7. In some embodiments, Form B has athermo-gravimetric analysis (TGA) thermogram substantially similar tothe one set forth in FIG. 8. In some embodiments, Form B has a DSCthermogram with an endotherm having an onset at about 99-106° C. and apeak at about 115-118° C.

In some embodiments, Form B has an observed aqueous solubility of about0.0096 mg/mL at a pH of about 7.42.

In some embodiments, Form B was obtained from a mixture of methanol andwater.

In some embodiments, Form B is unsolvated. In some embodiments, Form Bis anhydrous.

Compound 1, Form C

In some embodiments, Compound 1 is crystalline. In some embodiments,Compound 1 is crystalline Form C. Crystalline Form C of Compound 1 ischaracterized as having at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 9;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 7.0±0.1° 2-Theta, 14.0±0.1° 2-Theta,        15.7±0.1° 2-Theta, 18.2±0.1° 2-Theta, 19.1±0.1° 2-Theta,        19.5±0.1° 2-Theta, 20.3±0.1° 2-Theta, 22.1±0.1° 2-Theta, and        22.9±0.1° 2-Theta;    -   (c) a DSC thermogram substantially similar to the one set forth        in FIG. 10;    -   (d) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 11;    -   (e) a DSC thermogram with an endotherm having an onset at about        134-135° C. and a peak at about 137-139° C.;    -   or    -   (f) combinations thereof.

In some embodiments, Form C of Compound 1 is characterized as having atleast two of the properties selected from (a) to (e). In someembodiments, Form C of Compound 1 is characterized as having at leastthree of the properties selected from (a) to (e). In some embodiments,Form C of Compound 1 is characterized as having at least four of theproperties selected from (a) to (e). In some embodiments, Form C ofCompound 1 is characterized as having properties (a) to (e).

In some embodiments, Form C has an X-Ray powder diffraction (XRPD)pattern substantially the same as shown in FIG. 9. In some embodiments,Form C has an X-ray powder diffraction (XRPD) pattern withcharacteristic peaks at 7.0±0.1° 2-Theta, 14.0±0.1° 2-Theta, 15.7±0.1°2-Theta, 18.2±0.1° 2-Theta, 19.1±0.1° 2-Theta, 19.5±0.1° 2-Theta,20.3±0.1° 2-Theta, 22.1±0.1° 2-Theta, and 22.9±0.1° 2-Theta.

In some embodiments, Form C has a DSC thermogram substantially similarto the one set forth in FIG. 10. In some embodiments, Form C has athermo-gravimetric analysis (TGA) thermogram substantially similar tothe one set forth in FIG. 11. In some embodiments, Form C has a DSCthermogram with an endotherm having an onset at about 134-135° C. and apeak at about 137-139° C.

In some embodiments, Form C was obtained from a mixture of methanol andwater. In some embodiments, Form C was obtained from methanol.

In some embodiments, Form C is unsolvated. In some embodiments, Form Cis anhydrous.

Compound 1, Form D

In some embodiments, Compound 1 is crystalline. In some embodiments,Compound 1 is crystalline Form D. Crystalline Form D of Compound 1 ischaracterized as having at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 12;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 7.2±0.1° 2-Theta, 8.0±0.1° 2-Theta,        9.2±0.1° 2-Theta, 14.5±0.1° 2-Theta, 18.5±0.1° 2-Theta,        19.5±0.1° 2-Theta, 20.7±0.1° 2-Theta, 21.0±0.1° 2-Theta,        21.9±0.1° 2-Theta, and 22.4±0.1° 2-Theta;    -   (c) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 13;    -   or    -   (d) combinations thereof.

In some embodiments, Form D of Compound 1 is characterized as having atleast two of the properties selected from (a) to (c). In someembodiments, Form D of Compound 1 is characterized as having properties(a), (b), and (c).

In some embodiments, Form D has an X-Ray powder diffraction (XRPD)pattern substantially the same as shown in FIG. 12. In some embodiments,Form D has an X-ray powder diffraction (XRPD) pattern withcharacteristic peaks at 7.2±0.1° 2-Theta, 8.0±0.1° 2-Theta, 9.2±0.1°2-Theta, 14.5±0.1° 2-Theta, 18.5±0.1° 2-Theta, 19.5±0.1° 2-Theta,20.7±0.1° 2-Theta, 21.0±0.1° 2-Theta, 21.9±0.1° 2-Theta, and 22.4±0.1°2-Theta.

In some embodiments, Form D has a thermo-gravimetric analysis (TGA)thermogram substantially similar to the one set forth in FIG. 13.

In some embodiments, Form D was obtained from methyl isobutyl ketone(MIBK). In some embodiments, Form D is solvated. In some embodiments,Form D is solvated with methyl isobutyl ketone (MIBK).

Compound 1, Form E

In some embodiments, Compound 1 is crystalline. In some embodiments,Compound 1 is crystalline Form E. Crystalline Form E of Compound 1 ischaracterized as having at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 14;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 7.8±0.1° 2-Theta, 8.8±0.1° 2-Theta,        16.1±0.1° 2-Theta, 18.1±0.1° 2-Theta, 19.3±0.1° 2-Theta,        19.5±0.1° 2-Theta, 20.5±0.1° 2-Theta, 21.6±0.1° 2-Theta, and        25.2±0.1° 2-Theta;    -   (c) a DSC thermogram substantially similar to the one set forth        in FIG. 15;    -   (d) a thermo-gravimetric analysis (TGA) thermogram substantially        similar to the one set forth in FIG. 15;    -   or    -   (e) combinations thereof.

In some embodiments, Form E of Compound 1 is characterized as having atleast two of the properties selected from (a) to (d). In someembodiments, Form E of Compound 1 is characterized as having at leastthree of the properties selected from (a) to (d). In some embodiments,Form E of Compound 1 is characterized as having properties (a) to (d).

In some embodiments, Form E has an X-Ray powder diffraction (XRPD)pattern substantially the same as shown in FIG. 14. In some embodiments,Form E has an X-ray powder diffraction (XRPD) pattern withcharacteristic peaks at 7.8±0.1° 2-Theta, 8.8±0.1° 2-Theta, 16.1±0.1°2-Theta, 18.1±0.1° 2-Theta, 19.3±0.1° 2-Theta, 19.5±0.1° 2-Theta,20.5±0.1° 2-Theta, 21.6±0.1° 2-Theta, and 25.2±0.1° 2-Theta.

In some embodiments, Form E has a DSC thermogram substantially similarto the one set forth in FIG. 15. In some embodiments, Form E has athermo-gravimetric analysis (TGA) thermogram substantially similar tothe one set forth in FIG. 15.

In some embodiments, Form E was obtained from toluene.

In some embodiments, Form E is solvated. In some embodiments, Form E issolvated with toluene.

Compound 1, Form F

In some embodiments, Compound 1 is crystalline. In some embodiments,Compound 1 is crystalline Form F. Crystalline Form F of Compound 1 ischaracterized as having at least one of the following properties:

-   -   (a) an X-Ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 16;    -   (b) an X-ray powder diffraction (XRPD) pattern with        characteristic peaks at 6.2±0.1° 2-Theta, 10.1±0.1° 2-Theta,        17.6±0.1° 2-Theta, 18.6±0.1° 2-Theta, 20.0±0.1° 2-Theta,        20.4±0.1° 2-Theta, 20.7±0.1° 2-Theta, 22.4±0.1° 2-Theta,        23.0±0.1° 2-Theta, 23.2±0.1° 2-Theta, 24.4±0.1° 2-Theta,        25.1±0.1° 2-Theta, 27.6±0.1° 2-Theta, 29.3±0.1° 2-Theta, and        29.7±0.1° 2-Theta;    -   (c unit cell parameters substantially equal to the following at        1002K:

Crystal system Triclinic Space group P1 a 9.6332(3) Å α 105.762(3)° b9.7536(4) Å β  95.132(2)° c 15.0592(4) Å  γ 111.332(3)° V 1240.15(7) Å³Z 1 Density 1.308 Mg/m³ (calculated) Absorption 0.726 mm⁻¹ coefficientWavelength 1.54178 Å F(000) 518

-   -   or    -   (d) combinations thereof.

In some embodiments, Form F of Compound 1 is characterized as having atleast two of the properties selected from (a) to (c). In someembodiments, Form F of Compound 1 is characterized as having properties(a), (a), and (c).

In some embodiments, Form F has an X-Ray powder diffraction (XRPD)pattern substantially the same as shown in FIG. 16. In some embodiments,Form F has an X-ray powder diffraction (XRPD) pattern withcharacteristic peaks at 6.2±0.1° 2-Theta, 10.1±0.1° 2-Theta, 17.6±0.1°2-Theta, 18.6±0.1° 2-Theta, 20.0±0.1° 2-Theta, 20.4±0.1° 2-Theta,20.7±0.1° 2-Theta, 22.4±0.1° 2-Theta, 23.0±0.1° 2-Theta, 23.2±0.1°2-Theta, 24.4±0.1° 2-Theta, 25.1±0.1° 2-Theta, 27.6±0.1° 2-Theta,29.3±0.1° 2-Theta, and 29.7±0.1° 2-Theta.

In some embodiments, Form F has unit cell parameters substantially equalto the following at 100(2) K:

Crystal system Triclinic Space group P1 a 9.6332(3) Å α 105.762(3)° b9.7536(4) Å β  95.132(2)° c 15.0592(4) Å  γ 111.332(3)° V 1240.15(7) Å³Z 1 Density 1.308 Mg/m³ (calculated) Absorption 0.726 mm⁻¹ coefficientWavelength 1.54178 Å F(000) 518

In some embodiments, Form F was obtained from methanol.

In some embodiments, Form F is solvated. In some embodiments, Form F issolvated with methanol.

Preparation of Crystalline Forms

In some embodiments, crystalline forms of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneare prepared as outlined in the Examples. It is noted that solvents,temperatures and other reaction conditions presented herein may vary.

Suitable Solvents

Therapeutic agents that are administrable to mammals, such as humans,must be prepared by following regulatory guidelines. Such governmentregulated guidelines are referred to as Good Manufacturing Practice(GMP). GMP guidelines outline acceptable contamination levels of activetherapeutic agents, such as, for example, the amount of residual solventin the final product. Preferred solvents are those that are suitable foruse in GMP facilities and consistent with industrial safety concerns.Categories of solvents are defined in, for example, the InternationalConference on Harmonization of Technical Requirements for Registrationof Pharmaceuticals for Human Use (ICH), “Impurities: Guidelines forResidual Solvents, Q3C(R3), (November 2005).

Solvents are categorized into three classes. Class 1 solvents are toxicand are to be avoided. Class 2 solvents are solvents to be limited inuse during the manufacture of the therapeutic agent. Class 3 solventsare solvents with low toxic potential and of lower risk to human health.Data for Class 3 solvents indicate that they are less toxic in acute orshort-term studies and negative in genotoxicity studies.

Class 1 solvents, which are to be avoided, include: benzene; carbontetrachloride; 1,2-dichloroethane; 1,1-dichloroethene; and1,1,1-trichloroethane.

Examples of Class 2 solvents are: acetonitrile, chlorobenzene,chloroform, cyclohexane, 1,2-dichloroethene, dichloromethane,1,2-dimethoxyethane, N,N-dimethylacetamide, N,N-dimethylformamide,1,4-dioxane, 2-ethoxyethanol, ethyleneglycol, formamide, hexane,methanol, 2-methoxyethanol, methylbutyl ketone, methylcyclohexane,N-methylpyrrolidine, nitromethane, pyridine, sulfolane, tetralin,toluene, 1,1,2-trichloroethene and xylene.

Class 3 solvents, which possess low toxicity, include: acetic acid,acetone, anisole, 1-butanol, 2-butanol, butyl acetate, tert-butylmethylether (MTBE), cumene, dimethyl sulfoxide, ethanol, ethyl acetate, ethylether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropylacetate, methyl acetate, 3-methyl-1-butanol, methylethyl ketone,methylisobutyl ketone, 2-methyl-1-propanol, pentane, 1-pentanol,1-propanol, 2-propanol, propyl acetate, and tetrahydrofuran.

Residual solvents in active pharmaceutical ingredients (APIs) originatefrom the manufacture of API. In some cases, the solvents are notcompletely removed by practical manufacturing techniques. Appropriateselection of the solvent for the synthesis of APIs may enhance theyield, or determine characteristics such as crystal form, purity, andsolubility. Therefore, the solvent is a critical parameter in thesynthetic process.

In some embodiments, compositions comprising Compound 1 comprise anorganic solvent(s). In some embodiments, compositions comprisingCompound 1 comprise a residual amount of an organic solvent(s). In someembodiments, compositions comprising Compound 1 comprise a residualamount of a Class 3 solvent. In some embodiments, the organic solvent isa Class 3 solvent. In some embodiments, the Class 3 solvent is selectedfrom the group consisting of acetic acid, acetone, anisole, 1-butanol,2-butanol, butyl acetate, tert-butylmethyl ether, cumene, dimethylsulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl formate, formicacid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate,3-methyl-1-butanol, methylethyl ketone, methylisobutyl ketone,2-methyl-1-propanol, pentane, 1-pentanol, 1-propanol, 2-propanol, propylacetate, and tetrahydrofuran. In some embodiments, the Class 3 solventis selected from ethyl acetate, isopropyl acetate,tert-butylmethylether, heptane, isopropanol, and ethanol.

Certain Terminology

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which the claimed subject matter belongs. It is to be understoodthat the foregoing general description and the following detaileddescription are exemplary and explanatory only and are not restrictiveof any subject matter claimed. In this application, the use of thesingular includes the plural unless specifically stated otherwise. Itmust be noted that, as used in the specification and the appendedclaims, the singular forms “a,” “an” and “the” include plural referentsunless the context clearly dictates otherwise. In this application, theuse of “or” means “and/or” unless stated otherwise. Furthermore, use ofthe term “including” as well as other forms, such as “include”,“includes,” and “included,” is not limiting.

The section headings used herein are for organizational purposes onlyand are not to be construed as limiting the subject matter described.All documents, or portions of documents, cited in the applicationincluding, but not limited to, patents, patent applications, articles,books, manuals, and treatises are hereby expressly incorporated byreference in their entirety for any purpose.

The term “acceptable” or “pharmaceutically acceptable”, with respect toa formulation, composition or ingredient, as used herein, means havingno persistent detrimental effect on the general health of the subjectbeing treated or does not abrogate the biological activity or propertiesof the compound, and is relatively nontoxic.

As used herein, the term “agonist” refers to a compound, the presence ofwhich results in a biological activity of a protein that is the same asthe biological activity resulting from the presence of a naturallyoccurring ligand for the protein, such as, for example, Btk.

As used herein, the term “partial agonist” refers to a compound thepresence of which results in a biological activity of a protein that isof the same type as that resulting from the presence of a naturallyoccurring ligand for the protein, but of a lower magnitude.

As used herein, the term “antagonist” refers to a compound, the presenceof which results in a decrease in the magnitude of a biological activityof a protein. In certain embodiments, the presence of an antagonistresults in complete inhibition of a biological activity of a protein,such as, for example, Btk. In certain embodiments, an antagonist is aninhibitor.

As used herein, “amelioration” of the symptoms of a particular disease,disorder or condition by administration of a particular compound orpharmaceutical composition refers to any lessening of severity, delay inonset, slowing of progression, or shortening of duration, whetherpermanent or temporary, lasting or transient that can be attributed toor associated with administration of the compound or composition.

“Bioavailability” refers to the percentage of Compound 1 dosed that isdelivered into the general circulation of the animal or human beingstudied. The total exposure (AUC_((0-∞)) of a drug when administeredintravenously is usually defined as 100% bioavailable (F %). “Oralbioavailability” refers to the extent to which Compound 1 is absorbedinto the general circulation when the pharmaceutical composition istaken orally as compared to intravenous injection.

“Blood plasma concentration” refers to the concentration of Compound 1in the plasma component of blood of a subject. It is understood that theplasma concentration of Compound 1 may vary significantly betweensubjects, due to variability with respect to metabolism and/or possibleinteractions with other therapeutic agents. In accordance with oneembodiment disclosed herein, the blood plasma concentration of Compound1 may vary from subject to subject. Likewise, values such as maximumplasma concentration (C_(max)) or time to reach maximum plasmaconcentration (T_(max)), or total area under the plasma concentrationtime curve (AUC_((0-∞)) may vary from subject to subject. Due to thisvariability, the amount necessary to constitute “a therapeuticallyeffective amount” of Compound 1 may vary from subject to subject.

The term “Bruton's tyrosine kinase,” as used herein, refers to Bruton'styrosine kinase from Homo sapiens, as disclosed in, e.g., U.S. Pat. No.6,326,469 (GenBank Accession No. NP_000052).

The term “Bruton's tyrosine kinase homolog,” as used herein, refers toorthologs of Bruton's tyrosine kinase, e.g., the orthologs from mouse(GenBank Acession No. AAB47246), dog (GenBank Acession No. XP_549139.),rat (GenBank Acession No. NP_001007799), chicken (GenBank Acession No.NP_989564), or zebra fish (GenBank Acession No. XP_698117), and fusionproteins of any of the foregoing that exhibit kinase activity towardsone or more substrates of Bruton's tyrosine kinase (e.g. a peptidesubstrate having the amino acid sequence “AVLESEEELYSSARQ”).

The terms “co-administration” or the like, as used herein, are meant toencompass administration of the selected therapeutic agents to a singlepatient, and are intended to include treatment regimens in which theagents are administered by the same or different route of administrationor at the same or different time.

The terms “effective amount” or “therapeutically effective amount,” asused herein, refer to a sufficient amount of an agent or a compoundbeing administered which will relieve to some extent one or more of thesymptoms of the disease or condition being treated. The result can bereduction and/or alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. Forexample, an “effective amount” for therapeutic uses is the amount of thecomposition including a compound as disclosed herein required to providea clinically significant decrease in disease symptoms without undueadverse side effects.

An appropriate “effective amount” in any individual case may bedetermined using techniques, such as a dose escalation study. The term“therapeutically effective amount” includes, for example, aprophylactically effective amount. An “effective amount” of a compounddisclosed herein is an amount effective to achieve a desiredpharmacologic effect or therapeutic improvement without undue adverseside effects. It is understood that “an effect amount” or “atherapeutically effective amount” can vary from subject to subject, dueto variation in metabolism of Compound 1, age, weight, general conditionof the subject, the condition being treated, the severity of thecondition being treated, and the judgment of the prescribing physician.By way of example only, therapeutically effective amounts may bedetermined by routine experimentation, including but not limited to adose escalation clinical trial.

The terms “enhance” or “enhancing” means to increase or prolong eitherin potency or duration a desired effect. By way of example, “enhancing”the effect of therapeutic agents refers to the ability to increase orprolong, either in potency or duration, the effect of therapeutic agentson during treatment of a disease, disorder or condition. An“enhancing-effective amount,” as used herein, refers to an amountadequate to enhance the effect of a therapeutic agent in the treatmentof a disease, disorder or condition. When used in a patient, amountseffective for this use will depend on the severity and course of thedisease, disorder or condition, previous therapy, the patient's healthstatus and response to the drugs, and the judgment of the treatingphysician.

The term “homologous cysteine,” as used herein refers to a cysteineresidue found with in a sequence position that is homologous to that ofcysteine 481 of Bruton's tyrosine kinase, as defined herein. Forexample, cysteine 482 is the homologous cysteine of the rat ortholog ofBruton's tyrosine kinase; cysteine 479 is the homologous cysteine of thechicken ortholog; and cysteine 481 is the homologous cysteine in thezebra fish ortholog. In another example, the homologous cysteine of TXK,a Tec kinase family member related to Bruton's tyrosine, is Cys 350.Other examples of kinases having homologous cysteines are shown inFIG. 1. See also the sequence alignments of tyrosine kinases (TK)published on the world wide web atkinase.com/human/kinome/phylogeny.html.

The term “identical,” as used herein, refers to two or more sequences orsubsequences which are the same. In addition, the term “substantiallyidentical,” as used herein, refers to two or more sequences which have apercentage of sequential units which are the same when compared andaligned for maximum correspondence over a comparison window, ordesignated region as measured using comparison algorithms or by manualalignment and visual inspection. By way of example only, two or moresequences may be “substantially identical” if the sequential units areabout 60% identical, about 65% identical, about 70% identical, about 75%identical, about 80% identical, about 85% identical, about 90%identical, or about 95% identical over a specified region. Suchpercentages to describe the “percent identity” of two or more sequences.The identity of a sequence can exist over a region that is at leastabout 75-100 sequential units in length, over a region that is about 50sequential units in length, or, where not specified, across the entiresequence. This definition also refers to the complement of a testsequence. By way of example only, two or more polypeptide sequences areidentical when the amino acid residues are the same, while two or morepolypeptide sequences are “substantially identical” if the amino acidresidues are about 60% identical, about 65% identical, about 70%identical, about 75% identical, about 80% identical, about 85%identical, about 90% identical, or about 95% identical over a specifiedregion. The identity can exist over a region that is at least about75-100 amino acids in length, over a region that is about 50 amino acidsin length, or, where not specified, across the entire sequence of apolypeptide sequence. In addition, by way of example only, two or morepolynucleotide sequences are identical when the nucleic acid residuesare the same, while two or more polynucleotide sequences are“substantially identical” if the nucleic acid residues are about 60%identical, about 65% identical, about 70% identical, about 75%identical, about 80% identical, about 85% identical, about 90%identical, or about 95% identical over a specified region. The identitycan exist over a region that is at least about 75-100 nucleic acids inlength, over a region that is about 50 nucleic acids in length, or,where not specified, across the entire sequence of a polynucleotidesequence.

The terms “inhibits”, “inhibiting”, or “inhibitor” of a kinase, as usedherein, refer to inhibition of enzymatic phosphotransferase activity.

The term “irreversible inhibitor,” as used herein, refers to a compoundthat, upon contact with a target protein (e.g., a kinase) causes theformation of a new covalent bond with or within the protein, whereby oneor more of the target protein's biological activities (e.g.,phosphotransferase activity) is diminished or abolished notwithstandingthe subsequent presence or absence of the irreversible inhibitor.

The term “irreversible Btk inhibitor,” as used herein, refers to aninhibitor of Btk that can form a covalent bond with an amino acidresidue of Btk. In one embodiment, the irreversible inhibitor of Btk canform a covalent bond with a Cys residue of Btk; in particularembodiments, the irreversible inhibitor can form a covalent bond with aCys 481 residue (or a homolog thereof) of Btk or a cysteine residue inthe homologous corresponding position of another tyrosine kinase.

The term “isolated,” as used herein, refers to separating and removing acomponent of interest from components not of interest. Isolatedsubstances can be in either a dry or semi-dry state, or in solution,including but not limited to an aqueous solution. The isolated componentcan be in a homogeneous state or the isolated component can be a part ofa pharmaceutical composition that comprises additional pharmaceuticallyacceptable carriers and/or excipients. By way of example only, nucleicacids or proteins are “isolated” when such nucleic acids or proteins arefree of at least some of the cellular components with which it isassociated in the natural state, or that the nucleic acid or protein hasbeen concentrated to a level greater than the concentration of its invivo or in vitro production. Also, by way of example, a gene is isolatedwhen separated from open reading frames which flank the gene and encodea protein other than the gene of interest.

The term “modulate,” as used herein, means to interact with a targeteither directly or indirectly so as to alter the activity of the target,including, by way of example only, to enhance the activity of thetarget, to inhibit the activity of the target, to limit the activity ofthe target, or to extend the activity of the target.

As used herein, the term “modulator” refers to a compound that alters anactivity of a molecule. For example, a modulator can cause an increaseor decrease in the magnitude of a certain activity of a moleculecompared to the magnitude of the activity in the absence of themodulator. In certain embodiments, a modulator is an inhibitor, whichdecreases the magnitude of one or more activities of a molecule. Incertain embodiments, an inhibitor completely prevents one or moreactivities of a molecule. In certain embodiments, a modulator is anactivator, which increases the magnitude of at least one activity of amolecule. In certain embodiments the presence of a modulator results inan activity that does not occur in the absence of the modulator.

The term “prophylactically effective amount,” as used herein, refersthat amount of a composition applied to a patient which will relieve tosome extent one or more of the symptoms of a disease, condition ordisorder being treated. In such prophylactic applications, such amountsmay depend on the patient's state of health, weight, and the like. It isconsidered well within the skill of the art for one to determine suchprophylactically effective amounts by routine experimentation,including, but not limited to, a dose escalation clinical trial.

The term “subject” as used herein, refers to an animal which is theobject of treatment, observation or experiment. By way of example only,a subject may be, but is not limited to, a mammal including, but notlimited to, a human.

As used herein, the term “target activity” refers to a biologicalactivity capable of being modulated by a selective modulator. Certainexemplary target activities include, but are not limited to, bindingaffinity, signal transduction, enzymatic activity, tumor growth,inflammation or inflammation-related processes, and amelioration of oneor more symptoms associated with a disease or condition.

As used herein, the term “target protein” refers to a molecule or aportion of a protein capable of being bound by a selective bindingcompound. In certain embodiments, a target protein is Btk.

The terms “treat,” “treating” or “treatment”, as used herein, includealleviating, abating or ameliorating a disease or condition symptoms,preventing additional symptoms, ameliorating or preventing theunderlying metabolic causes of symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition. The terms “treat,”“treating” or “treatment”, include, but are not limited to, prophylacticand/or therapeutic treatments.

As used herein, the IC₅₀ refers to an amount, concentration or dosage ofa particular test compound that achieves a 50% inhibition of a maximalresponse, such as inhibition of Btk, in an assay that measures suchresponse.

As used herein, EC₅₀ refers to a dosage, concentration or amount of aparticular test compound that elicits a dose-dependent response at 50%of maximal expression of a particular response that is induced, provokedor potentiated by the particular test compound.

Pharmaceutical Compositions/Formulations

Pharmaceutical compositions may be formulated in a conventional mannerusing one or more physiologically acceptable carriers includingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically.

Proper formulation is dependent upon the route of administration chosen.Any of the well-known techniques, carriers, and excipients may be usedas suitable and as understood in the art.

A summary of pharmaceutical compositions described herein may be found,for example, in Remington: The Science and Practice of Pharmacy,Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, JohnE., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton,Pa. 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins 1999), herein incorporated by reference in their entirety.

A pharmaceutical composition, as used herein, refers to a mixture ofCompound 1 with other chemical components, such as carriers,stabilizers, diluents, dispersing agents, suspending agents, thickeningagents, and/or excipients. The pharmaceutical composition facilitatesadministration of the compound to a mammal. In practicing the methods oftreatment or use provided herein, therapeutically effective amounts ofCompound 1 are administered in a pharmaceutical composition to a mammalhaving a disease, disorder, or condition to be treated. Preferably, themammal is a human. A therapeutically effective amount can vary widelydepending on the severity of the disease, the age and relative health ofthe subject, the potency of the compound used and other factors. Thecompounds can be used singly or in combination with one or moretherapeutic agents as components of mixtures.

The term “pharmaceutical combination” as used herein, means a productthat results from the mixing or combining of more than one activeingredient and includes both fixed and non-fixed combinations of theactive ingredients. The term “fixed combination” means that the activeingredients, e.g. Compound 1 and a co-agent, are both administered to apatient simultaneously in the form of a single entity or dosage. Theterm “non-fixed combination” means that the active ingredients, e.g.Compound 1 and a co-agent, are administered to a patient as separateentities either simultaneously, concurrently or sequentially with nospecific intervening time limits, wherein such administration provideseffective levels of the two compounds in the body of the patient. Thelatter also applies to cocktail therapy, e.g. the administration ofthree or more active ingredients.

In some embodiments, crystalline Compound 1 is incorporated intopharmaceutical compositions to provide solid oral dosage forms. In otherembodiments, crystalline Compound 1 is used to prepare pharmaceuticalcompositions other than oral solid dosage forms. The pharmaceuticalformulations described herein can be administered to a subject bymultiple administration routes, including but not limited to, oral,parenteral (e.g., intravenous, subcutaneous, intramuscular), intranasal,buccal, topical, rectal, or transdermal administration routes. Thepharmaceutical formulations described herein include, but are notlimited to, aqueous liquid dispersions, self-emulsifying dispersions,solid solutions, liposomal dispersions, aerosols, solid dosage forms,powders, immediate release formulations, controlled releaseformulations, fast melt formulations, tablets, capsules, pills, delayedrelease formulations, extended release formulations, pulsatile releaseformulations, multiparticulate formulations, and mixed immediate andcontrolled release formulations.

Pharmaceutical compositions including a compound described herein may bemanufactured in a conventional manner, such as, by way of example only,by means of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or compressionprocesses.

Dosage Forms

The pharmaceutical compositions described herein can be formulated foradministration to a mammal via any conventional means including, but notlimited to, oral, parenteral (e.g., intravenous, subcutaneous, orintramuscular), buccal, intranasal, rectal or transdermal administrationroutes. As used herein, the term “subject” is used to mean an animal,preferably a mammal, including a human or non-human. The terms patientand subject may be used interchangeably.

Moreover, the pharmaceutical compositions described herein, whichinclude Compound 1 can be formulated into any suitable dosage form,including but not limited to, solid oral dosage forms, controlledrelease formulations, fast melt formulations, effervescent formulations,tablets, powders, pills, capsules, delayed release formulations,extended release formulations, pulsatile release formulations,multiparticulate formulations, and mixed immediate release andcontrolled release formulations.

Pharmaceutical preparations for oral use can be obtained by mixing oneor more solid excipient with one or more of the compounds describedherein, optionally grinding the resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Suitable excipients include, forexample, fillers such as sugars, including lactose, sucrose, mannitol,or sorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methylcellulose, microcrystalline cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or otherssuch as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. Ifdesired, disintegrating agents may be added, such as the cross-linkedcroscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or asalt thereof such as sodium alginate.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added. All formulations fororal administration should be in dosages suitable for suchadministration.

In some embodiments, the solid dosage forms disclosed herein may be inthe form of a tablet, (including a suspension tablet, a fast-melttablet, a bite-disintegration tablet, a rapid-disintegration tablet, aneffervescent tablet, or a caplet), a pill, a powder (including a sterilepackaged powder, a dispensable powder, or an effervescent powder) acapsule (including both soft or hard capsules, e.g., capsules made fromanimal-derived gelatin or plant-derived HPMC, or “sprinkle capsules”),solid dispersion, solid solution, bioerodible dosage form, controlledrelease formulations, pulsatile release dosage forms, multiparticulatedosage forms, pellets, granules, or an aerosol. In other embodiments,the pharmaceutical formulation is in the form of a powder. In stillother embodiments, the pharmaceutical formulation is in the form of atablet, including but not limited to, a fast-melt tablet. Additionally,pharmaceutical formulations described herein may be administered as asingle capsule or in multiple capsule dosage form. In some embodiments,the pharmaceutical formulation is administered in two, or three, orfour, capsules or tablets.

In some embodiments, solid dosage forms, e.g., tablets, effervescenttablets, and capsules, are prepared by mixing particles of Compound 1with one or more pharmaceutical excipients to form a bulk blendcomposition. When referring to these bulk blend compositions ashomogeneous, it is meant that the particles of Compound 1 are dispersedevenly throughout the composition so that the composition may be readilysubdivided into equally effective unit dosage forms, such as tablets,pills, and capsules. The individual unit dosages may also include filmcoatings, which disintegrate upon oral ingestion or upon contact withdiluent. These formulations can be manufactured by conventionalpharmacological techniques.

Conventional pharmacological techniques include, e.g., one or acombination of methods: (1) dry mixing, (2) direct compression, (3)milling, (4) dry or non-aqueous granulation, (5) wet granulation, or (6)fusion. See, e.g., Lachman et al., The Theory and Practice of IndustrialPharmacy (1986). Other methods include, e.g., spray drying, pan coating,melt granulation, granulation, fluidized bed spray drying or coating(e.g., wurster coating), tangential coating, top spraying, tableting,extruding and the like.

The pharmaceutical solid dosage forms described herein can includeCompound 1 and one or more pharmaceutically acceptable additives such asa compatible carrier, binder, filling agent, suspending agent, flavoringagent, sweetening agent, disintegrating agent, dispersing agent,surfactant, lubricant, colorant, diluent, solubilizer, moistening agent,plasticizer, stabilizer, penetration enhancer, wetting agent,anti-foaming agent, antioxidant, preservative, or one or morecombination thereof. In still other aspects, using standard coatingprocedures, such as those described in Remington's PharmaceuticalSciences, 20th Edition (2000), a film coating is provided around theformulation of Compound 1. In one embodiment, some or all of theparticles of the Compound 1 are coated. In another embodiment, some orall of the particles of the Compound 1 are microencapsulated. In stillanother embodiment, the particles of the Compound 1 are notmicroencapsulated and are uncoated.

Suitable carriers for use in the solid dosage forms described hereininclude, but are not limited to, acacia, gelatin, colloidal silicondioxide, calcium glycerophosphate, calcium lactate, maltodextrin,glycerine, magnesium silicate, sodium caseinate, soy lecithin, sodiumchloride, tricalcium phosphate, dipotassium phosphate, sodium stearoyllactylate, carrageenan, monoglyceride, diglyceride, pregelatinizedstarch, hydroxypropylmethylcellulose, hydroxypropylmethylcelluloseacetate stearate, sucrose, microcrystalline cellulose, lactose, mannitoland the like.

Suitable filling agents for use in the solid dosage forms describedherein include, but are not limited to, lactose, calcium carbonate,calcium phosphate, dibasic calcium phosphate, calcium sulfate,microcrystalline cellulose, cellulose powder, dextrose, dextrates,dextran, starches, pregelatinized starch, hydroxypropylmethycellulose(HPMC), hydroxypropylmethycellulose phthalate,hydroxypropylmethylcellulose acetate stearate (HPMCAS), sucrose,xylitol, lactitol, mannitol, sorbitol, sodium chloride, polyethyleneglycol, and the like.

In order to release the Compound 1 from a solid dosage form matrix asefficiently as possible, disintegrants are often used in theformulation, especially when the dosage forms are compressed withbinder. Disintegrants help rupturing the dosage form matrix by swellingor capillary action when moisture is absorbed into the dosage form.Suitable disintegrants for use in the solid dosage forms describedherein include, but are not limited to, natural starch such as cornstarch or potato starch, a pregelatinized starch such as National 1551or Amijel®, or sodium starch glycolate such as Promogel® or Explotab®, acellulose such as a wood product, methylcrystalline cellulose, e.g.,Avicel®, Avicel® PH101, Avicel® PH102, Avicel® PH105, Elcema® P100,Emcocel®, Vivacel®, Ming Tia®, and Solka-Floc®, methylcellulose,croscarmellose, or a cross-linked cellulose, such as cross-linked sodiumcarboxymethylcellulose (Ac-Di-Sol©), cross-linkedcarboxymethylcellulose, or cross-linked croscarmellose, a cross-linkedstarch such as sodium starch glycolate, a cross-linked polymer such ascrospovidone, a cross-linked polyvinylpyrrolidone, alginate such asalginic acid or a salt of alginic acid such as sodium alginate, a claysuch as Veegum® HV (magnesium aluminum silicate), a gum such as agar,guar, locust bean, Karaya, pectin, or tragacanth, sodium starchglycolate, bentonite, a natural sponge, a surfactant, a resin such as acation-exchange resin, citrus pulp, sodium lauryl sulfate, sodium laurylsulfate in combination starch, and the like. In some embodimentsprovided herein, the disintegrating agent is selected from the groupconsisting of natural starch, a pregelatinized starch, a sodium starch,methylcrystalline cellulose, methylcellulose, croscarmellose,croscarmellose sodium, cross-linked sodium carboxymethylcellulose,cross-linked carboxymethylcellulose, cross-linked croscarmellose,cross-linked starch such as sodium starch glycolate, cross-linkedpolymer such as crospovidone, cross-linked polyvinylpyrrolidone, sodiumalginate, a clay, or a gum. In some embodiments provided herein, thedisintegrating agent is croscarmellose sodium.

Binders impart cohesiveness to solid oral dosage form formulations: forpowder filled capsule formulation, they aid in plug formation that canbe filled into soft or hard shell capsules and for tablet formulation,they ensure the tablet remaining intact after compression and helpassure blend uniformity prior to a compression or fill step. Materialssuitable for use as binders in the solid dosage forms described hereininclude, but are not limited to, carboxymethylcellulose, methylcellulose(e.g., Methocel®), hydroxypropylmethylcellulose (e.g. Hypromellose USPPharmacoat-603, hydroxypropylmethylcellulose acetate stearate (AqoateHS-LF and HS), hydroxyethylcellulose, hydroxypropylcellulose (e.g.,Klucel®), ethylcellulose (e.g., Ethocel®), and microcrystallinecellulose (e.g., Avicel®), microcrystalline dextrose, amylose, magnesiumaluminum silicate, polysaccharide acids, bentonites, gelatin,polyvinylpyrrolidone/vinyl acetate copolymer, crospovidone, povidone,starch, pregelatinized starch, tragacanth, dextrin, a sugar, such assucrose (e.g., Dipac®), glucose, dextrose, molasses, mannitol, sorbitol,xylitol (e.g., Xylitab®), lactose, a natural or synthetic gum such asacacia, tragacanth, ghatti gum, mucilage of isapol husks, starch,polyvinylpyrrolidone (e.g., Povidone® CL, Kollidon® CL, Polyplasdone®XL-10, and Povidone® K-12), larch arabogalactan, Veegum®, polyethyleneglycol, waxes, sodium alginate, and the like.

In general, binder levels of 20-70% are used in powder-filled gelatincapsule formulations. Binder usage level in tablet formulations varieswhether direct compression, wet granulation, roller compaction, or usageof other excipients such as fillers which itself can act as moderatebinder. Formulators skilled in art can determine the binder level forthe formulations, but binder usage level of up to 70% in tabletformulations is common.

Suitable lubricants or glidants for use in the solid dosage formsdescribed herein include, but are not limited to, stearic acid, calciumhydroxide, talc, corn starch, sodium stearyl fumerate, alkali-metal andalkaline earth metal salts, such as aluminum, calcium, magnesium, zinc,stearic acid, sodium stearates, magnesium stearate, zinc stearate,waxes, Stearowet®, boric acid, sodium benzoate, sodium acetate, sodiumchloride, leucine, a polyethylene glycol or a methoxypolyethylene glycolsuch as Carbowax™, PEG 4000, PEG 5000, PEG 6000, propylene glycol,sodium oleate, glyceryl behenate, glyceryl palmitostearate, glycerylbenzoate, magnesium or sodium lauryl sulfate, and the like. In someembodiments provided herein, the lubricant is selected from the groupconsisting of stearic acid, calcium hydroxide, talc, corn starch, sodiumstearyl fumerate, stearic acid, sodium stearates, magnesium stearate,zinc stearate, and waxes. In some embodiments provided herein, thelubricant is magnesium stearate.

Suitable diluents for use in the solid dosage forms described hereininclude, but are not limited to, sugars (including lactose, sucrose, anddextrose), polysaccharides (including dextrates and maltodextrin),polyols (including mannitol, xylitol, and sorbitol), cyclodextrins andthe like. In some embodiments provided herein, the diluent is selectedfrom the group consisting of lactose, sucrose, dextrose, dextrates,maltodextrin, mannitol, xylitol, sorbitol, cyclodextrins, calciumphosphate, calcium sulfate, starches, modified starches,microcrystalline cellulose, microcellulose, and talc. In someembodiments provided herein, the diluent is microcrystalline cellulose.

The term “non water-soluble diluent” represents compounds typically usedin the formulation of pharmaceuticals, such as calcium phosphate,calcium sulfate, starches, modified starches and microcrystallinecellulose, and microcellulose (e.g., having a density of about 0.45g/cm³, e.g. Avicel, powdered cellulose), and talc.

Suitable wetting agents for use in the solid dosage forms describedherein include, for example, oleic acid, glyceryl monostearate, sorbitanmonooleate, sorbitan monolaurate, triethanolamine oleate,polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitanmonolaurate, quaternary ammonium compounds (e.g., Polyquat 10®), sodiumoleate, sodium lauryl sulfate, magnesium stearate, sodium docusate,triacetin, vitamin E TPGS and the like.

Suitable surfactants for use in the solid dosage forms described hereininclude, for example, sodium lauryl sulfate, sorbitan monooleate,polyoxyethylene sorbitan monooleate, polysorbates, polaxomers, bilesalts, glyceryl monostearate, copolymers of ethylene oxide and propyleneoxide, e.g., Pluronic® (BASF), and the like. In some embodimentsprovided herein, the surfactant is selected from the group consisting ofsodium lauryl sulfate, sorbitan monooleate, polyoxyethylene sorbitanmonooleate, polysorbates, polaxomers, bile salts, glyceryl monostearate,copolymers of ethylene oxide and propylene oxide. In some embodimentsprovided herein, the surfactant is sodium lauryl sulfate.

Suitable suspending agents for use in the solid dosage forms describedhere include, but are not limited to, polyvinylpyrrolidone, e.g.,polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidoneK25, or polyvinylpyrrolidone K30, polyethylene glycol, e.g., thepolyethylene glycol can have a molecular weight of about 300 to about6000, or about 3350 to about 4000, or about 7000 to about 5400, vinylpyrrolidone/vinyl acetate copolymer (S630), sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,polysorbate-80, hydroxyethylcellulose, sodium alginate, gums, such as,e.g., gum tragacanth and gum acacia, guar gum, xanthans, includingxanthan gum, sugars, cellulosics, such as, e.g., sodiumcarboxymethylcellulose, methylcellulose, sodium carboxymethylcellulose,hydroxypropylmethylcellulose, hydroxyethylcellulose, polysorbate-80,sodium alginate, polyethoxylated sorbitan monolaurate, polyethoxylatedsorbitan monolaurate, povidone and the like.

Suitable antioxidants for use in the solid dosage forms described hereininclude, for example, e.g., butylated hydroxytoluene (BHT), sodiumascorbate, and tocopherol.

It should be appreciated that there is considerable overlap betweenadditives used in the solid dosage forms described herein. Thus, theabove-listed additives should be taken as merely exemplary, and notlimiting, of the types of additives that can be included in solid dosageforms described herein. The amounts of such additives can be readilydetermined by one skilled in the art, according to the particularproperties desired.

In other embodiments, one or more layers of the pharmaceuticalformulation are plasticized. Illustratively, a plasticizer is generallya high boiling point solid or liquid. Suitable plasticizers can be addedfrom about 0.01% to about 50% by weight (w/w) of the coatingcomposition. Plasticizers include, but are not limited to, diethylphthalate, citrate esters, polyethylene glycol, glycerol, acetylatedglycerides, triacetin, polypropylene glycol, polyethylene glycol,triethyl citrate, dibutyl sebacate, stearic acid, stearol, stearate, andcastor oil.

Compressed tablets are solid dosage forms prepared by compacting thebulk blend of the formulations described above. In various embodiments,compressed tablets which are designed to dissolve in the mouth willinclude one or more flavoring agents. In other embodiments, thecompressed tablets will include a film surrounding the final compressedtablet. In some embodiments, the film coating can provide a delayedrelease of Compound 1 from the formulation. In other embodiments, thefilm coating aids in patient compliance (e.g., Opadry® coatings or sugarcoating). Film coatings including Opadry® typically range from about 1%to about 3% of the tablet weight. In other embodiments, the compressedtablets include one or more excipients.

A capsule may be prepared, for example, by placing the bulk blend of theformulation of Compound 1 inside of a capsule. In some embodiments, theformulations (non-aqueous suspensions and solutions) are placed in asoft gelatin capsule. In other embodiments, the formulations are placedin standard gelatin capsules or non-gelatin capsules such as capsulescomprising HPMC. In other embodiments, the formulation is placed in asprinkle capsule, wherein the capsule may be swallowed whole or thecapsule may be opened and the contents sprinkled on food prior toeating. In some embodiments, the therapeutic dose is split into multiple(e.g., two, three, or four) capsules. In some embodiments, the entiredose of the formulation is delivered in a capsule form.

In various embodiments, the particles of Compound 1 and one or moreexcipients are dry blended and compressed into a mass, such as a tablet,having a hardness sufficient to provide a pharmaceutical compositionthat substantially disintegrates within less than about 30 minutes, lessthan about 35 minutes, less than about 40 minutes, less than about 45minutes, less than about 50 minutes, less than about 55 minutes, or lessthan about 60 minutes, after oral administration, thereby releasing theformulation into the gastrointestinal fluid.

In another aspect, dosage forms may include microencapsulatedformulations. In some embodiments, one or more other compatiblematerials are present in the microencapsulation material. Exemplarymaterials include, but are not limited to, pH modifiers, erosionfacilitators, anti-foaming agents, antioxidants, flavoring agents, andcarrier materials such as binders, suspending agents, disintegrationagents, filling agents, surfactants, solubilizers, stabilizers,lubricants, wetting agents, and diluents.

Materials useful for the microencapsulation described herein includematerials compatible with Compound 1 which sufficiently isolate theCompound 1 from other non-compatible excipients. Materials compatiblewith Compound 1 are those that delay the release of the compounds ofCompound 1 in vivo.

Exemplary microencapsulation materials useful for delaying the releaseof the formulations including compounds described herein, include, butare not limited to, hydroxypropyl cellulose ethers (HPC) such as Klucel®or Nisso HPC, low-substituted hydroxypropyl cellulose ethers (L-HPC),hydroxypropyl methyl cellulose ethers (HPMC) such as Seppifilm-LC,Pharmacoat®, Metolose SR, Methocel®-E, Opadry YS, PrimaFlo, BenecelMP824, and Benecel MP843, methylcellulose polymers such as Methocel®-A,hydroxypropylmethylcellulose acetate stearate Aqoat (HF-LS, HF-LG,HF-MS) and Metolose®, Ethylcelluloses (EC) and mixtures thereof such asE461, Ethocel®, Aqualon®-EC, Surelease®, Polyvinyl alcohol (PVA) such asOpadry AMB, hydroxyethylcelluloses such as Natrosol®,carboxymethylcelluloses and salts of carboxymethylcelluloses (CMC) suchas Aqualon®-CMC, polyvinyl alcohol and polyethylene glycol co-polymerssuch as Kollicoat IR®, monoglycerides (Myverol), triglycerides (KLX),polyethylene glycols, modified food starch, acrylic polymers andmixtures of acrylic polymers with cellulose ethers such as Eudragit®EPO, Eudragit® L30D-55, Eudragit® FS 30D Eudragit® L100-55, Eudragit®L100, Eudragit® 5100, Eudragit® RD100, Eudragit® E100, Eudragit® L12.5,Eudragit® S12.5, Eudragit® NE30D, and Eudragit® NE 40D, celluloseacetate phthalate, sepifilms such as mixtures of HPMC and stearic acid,cyclodextrins, and mixtures of these materials.

In still other embodiments, plasticizers such as polyethylene glycols,e.g., PEG 300, PEG 400, PEG 600, PEG 1450, PEG 3350, and PEG 800,stearic acid, propylene glycol, oleic acid, and triacetin areincorporated into the microencapsulation material. In other embodiments,the microencapsulating material useful for delaying the release of thepharmaceutical compositions is from the USP or the National Formulary(NF). In yet other embodiments, the microencapsulation material isKlucel. In still other embodiments, the microencapsulation material ismethocel.

Microencapsulated Compound 1 may be formulated by methods known by oneof ordinary skill in the art. Such known methods include, e.g., spraydrying processes, spinning disk-solvent processes, hot melt processes,spray chilling methods, fluidized bed, electrostatic deposition,centrifugal extrusion, rotational suspension separation, polymerizationat liquid-gas or solid-gas interface, pressure extrusion, or sprayingsolvent extraction bath. In addition to these, several chemicaltechniques, e.g., complex coacervation, solvent evaporation,polymer-polymer incompatibility, interfacial polymerization in liquidmedia, in situ polymerization, in-liquid drying, and desolvation inliquid media could also be used. Furthermore, other methods such asroller compaction, extrusion/spheronization, coacervation, ornanoparticle coating may also be used.

In one embodiment, the particles of Compound 1 are microencapsulatedprior to being formulated into one of the above forms. In still anotherembodiment, some or most of the particles are coated prior to beingfurther formulated by using standard coating procedures, such as thosedescribed in Remington's Pharmaceutical Sciences, 20th Edition (2000).

In other embodiments, the solid dosage formulations of the Compound 1are plasticized (coated) with one or more layers. Illustratively, aplasticizer is generally a high boiling point solid or liquid. Suitableplasticizers can be added from about 0.01% to about 50% by weight (w/w)of the coating composition. Plasticizers include, but are not limitedto, diethyl phthalate, citrate esters, polyethylene glycol, glycerol,acetylated glycerides, triacetin, polypropylene glycol, polyethyleneglycol, triethyl citrate, dibutyl sebacate, stearic acid, stearol,stearate, and castor oil.

In other embodiments, a powder including the formulations with Compound1 may be formulated to include one or more pharmaceutical excipients andflavors. Such a powder may be prepared, for example, by mixing theformulation and optional pharmaceutical excipients to form a bulk blendcomposition. Additional embodiments also include a suspending agentand/or a wetting agent. This bulk blend is uniformly subdivided intounit dosage packaging or multi-dosage packaging units.

In still other embodiments, effervescent powders are also prepared inaccordance with the present disclosure. Effervescent salts have beenused to disperse medicines in water for oral administration.Effervescent salts are granules or coarse powders containing a medicinalagent in a dry mixture, usually composed of sodium bicarbonate, citricacid and/or tartaric acid. When salts of the compositions describedherein are added to water, the acids and the base react to liberatecarbon dioxide gas, thereby causing “effervescence.” Examples ofeffervescent salts include, e.g., the following ingredients: sodiumbicarbonate or a mixture of sodium bicarbonate and sodium carbonate,citric acid and/or tartaric acid. Any acid-base combination that resultsin the liberation of carbon dioxide can be used in place of thecombination of sodium bicarbonate and citric and tartaric acids, as longas the ingredients were suitable for pharmaceutical use and result in apH of about 6.0 or higher.

In some embodiments, the solid dosage forms described herein can beformulated as enteric coated delayed release oral dosage forms, i.e., asan oral dosage form of a pharmaceutical composition as described hereinwhich utilizes an enteric coating to affect release in the smallintestine of the gastrointestinal tract. The enteric coated dosage formmay be a compressed or molded or extruded tablet/mold (coated oruncoated) containing granules, powder, pellets, beads or particles ofthe active ingredient and/or other composition components, which arethemselves coated or uncoated. The enteric coated oral dosage form mayalso be a capsule (coated or uncoated) containing pellets, beads orgranules of the solid carrier or the composition, which are themselvescoated or uncoated.

The term “delayed release” as used herein refers to the delivery so thatthe release can be accomplished at some generally predictable locationin the intestinal tract more distal to that which would have beenaccomplished if there had been no delayed release alterations. In someembodiments the method for delay of release is coating. Any coatingsshould be applied to a sufficient thickness such that the entire coatingdoes not dissolve in the gastrointestinal fluids at pH below about 5,but does dissolve at pH about 5 and above. It is expected that anyanionic polymer exhibiting a pH-dependent solubility profile can be usedas an enteric coating in the methods and compositions described hereinto achieve delivery to the lower gastrointestinal tract. In someembodiments the polymers described herein are anionic carboxylicpolymers. In other embodiments, the polymers and compatible mixturesthereof, and some of their properties, include, but are not limited to:

Shellac, also called purified lac, a refined product obtained from theresinous secretion of an insect. This coating dissolves in media ofpH>7;

Acrylic polymers. The performance of acrylic polymers (primarily theirsolubility in biological fluids) can vary based on the degree and typeof substitution. Examples of suitable acrylic polymers includemethacrylic acid copolymers and ammonium methacrylate copolymers. TheEudragit series E, L, S, RL, RS and NE (Rohm Pharma) are available assolubilized in organic solvent, aqueous dispersion, or dry powders. TheEudragit series RL, NE, and RS are insoluble in the gastrointestinaltract but are permeable and are used primarily for colonic targeting.The Eudragit series E dissolve in the stomach. The Eudragit series L,L-30D and S are insoluble in stomach and dissolve in the intestine;

Cellulose Derivatives. Examples of suitable cellulose derivatives are:ethyl cellulose; reaction mixtures of partial acetate esters ofcellulose with phthalic anhydride. The performance can vary based on thedegree and type of substitution. Cellulose acetate phthalate (CAP)dissolves in pH>6. Aquateric (FMC) is an aqueous based system and is aspray dried CAP psuedolatex with particles<1 μm. Other components inAquateric can include pluronics, Tweens, and acetylated monoglycerides.Other suitable cellulose derivatives include: cellulose acetatetrimellitate (Eastman); methylcellulose (Pharmacoat, Methocel);hydroxypropylmethyl cellulose phthalate (HPMCP); hydroxypropylmethylcellulose succinate (HPMCS); and hydroxypropylmethylcellulose acetatesuccinate (e.g., AQOAT (Shin Etsu)). The performance can vary based onthe degree and type of substitution. For example, HPMCP such as, HP-50,HP-55, HP-55S, HP-55F grades are suitable. The performance can varybased on the degree and type of substitution. For example, suitablegrades of hydroxypropylmethylcellulose acetate succinate include, butare not limited to, AS-LG (LF), which dissolves at pH 5, AS-MG (MF),which dissolves at pH 5.5, and AS-HG (HF), which dissolves at higher pH.These polymers are offered as granules, or as fine powders for aqueousdispersions; Poly Vinyl Acetate Phthalate (PVAP). PVAP dissolves inpH>5, and it is much less permeable to water vapor and gastric fluids.

In some embodiments, the coating can, and usually does, contain aplasticizer and possibly other coating excipients such as colorants,talc, and/or magnesium stearate, which are well known in the art.Suitable plasticizers include triethyl citrate (Citroflex 2), triacetin(glyceryl triacetate), acetyl triethyl citrate (Citroflec A2), Carbowax400 (polyethylene glycol 400), diethyl phthalate, tributyl citrate,acetylated monoglycerides, glycerol, fatty acid esters, propyleneglycol, and dibutyl phthalate. In particular, anionic carboxylic acrylicpolymers usually will contain 10-25% by weight of a plasticizer,especially dibutyl phthalate, polyethylene glycol, triethyl citrate andtriacetin. Conventional coating techniques such as spray or pan coatingare employed to apply coatings. The coating thickness must be sufficientto ensure that the oral dosage form remains intact until the desiredsite of topical delivery in the intestinal tract is reached.

Colorants, detackifiers, surfactants, antifoaming agents, lubricants(e.g., carnuba wax or PEG) may be added to the coatings besidesplasticizers to solubilize or disperse the coating material, and toimprove coating performance and the coated product.

In other embodiments, the formulations described herein, which includeCompound 1 are delivered using a pulsatile dosage form. A pulsatiledosage form is capable of providing one or more immediate release pulsesat predetermined time points after a controlled lag time or at specificsites. Many other types of controlled release systems known to those ofordinary skill in the art and are suitable for use with the formulationsdescribed herein. Examples of such delivery systems include, e.g.,polymer-based systems, such as polylactic and polyglycolic acid,plyanhydrides and polycaprolactone; porous matrices, nonpolymer-basedsystems that are lipids, including sterols, such as cholesterol,cholesterol esters and fatty acids, or neutral fats, such as mono-, di-and triglycerides; hydrogel release systems; silastic systems;peptide-based systems; wax coatings, bioerodible dosage forms,compressed tablets using conventional binders and the like. See, e.g.,Liberman et al., Pharmaceutical Dosage Forms, 2 Ed., Vol. 1, pp. 209-214(1990); Singh et al., Encyclopedia of Pharmaceutical Technology, 2^(nd)Ed., pp. 751-753 (2002); U.S. Pat. Nos. 4,327,725, 4,624,848, 4,968,509,5,461,140, 5,456,923, 5,516,527, 5,622,721, 5,686,105, 5,700,410,5,977,175, 6,465,014 and 6,932,983, each of which is specificallyincorporated by reference.

In some embodiments, pharmaceutical formulations are provided thatinclude particles of Compound 1 at least one dispersing agent orsuspending agent for oral administration to a subject. The formulationsmay be a powder and/or granules for suspension, and upon admixture withwater, a substantially uniform suspension is obtained.

It is to be appreciated that there is overlap between the above-listedadditives used in the aqueous dispersions or suspensions describedherein, since a given additive is often classified differently bydifferent practitioners in the field, or is commonly used for any ofseveral different functions. Thus, the above-listed additives should betaken as merely exemplary, and not limiting, of the types of additivesthat can be included in formulations described herein. The amounts ofsuch additives can be readily determined by one skilled in the art,according to the particular properties desired.

Dosing and Treatment Regimens

In some embodiments, the amount of Compound 1 that is administered to amammal is from 300 mg/day up to, and including, 1000 mg/day. In someembodiments, the amount of Compound 1 that is administered to a mammalis from 420 mg/day up to, and including, 840 mg/day. In someembodiments, the amount of Compound 1 that is administered to a mammalis about 420 mg/day, about 560 mg/day, or about 840 mg/day. In someembodiments, the amount of Compound 1 that is administered to a mammalis about 420 mg/day. In some embodiments, the amount of Compound 1 thatis administered to a mammal is about 560 mg/day. In some embodiments,the AUC₀₋₂₄ of Compound 1 is between about 150 and about 3500 ng*h/mL.In some embodiments, the AUC₀₋₂₄ of Compound 1 is between about 500 andabout 1100 ng*h/mL. In some embodiments, Compound 1 is administeredorally. In some embodiments, Compound 1 is administered once per day,twice per day, or three times per day. In some embodiments, Compound 1is administered daily. In some embodiments, Compound 1 is administeredonce daily. In some embodiments, Compound 1 is administered every otherday. In some embodiments, the Compound 1 is a maintenance therapy.

Compound 1 can be used in the preparation of medicaments for theinhibition of Btk or a homolog thereof, or for the treatment of diseasesor conditions that would benefit, at least in part, from inhibition ofBtk or a homolog thereof, including a subject diagnosed with ahematological malignancy. In addition, a method for treating any of thediseases or conditions described herein in a subject in need of suchtreatment, involves administration of pharmaceutical compositionscontaining Compound 1, or a pharmaceutically acceptable salt,pharmaceutically acceptable N-oxide, pharmaceutically active metabolite,pharmaceutically acceptable prodrug, or pharmaceutically acceptablesolvate thereof, in therapeutically effective amounts to said subject.

The compositions containing Compound 1 can be administered forprophylactic, therapeutic, or maintenance treatment. In someembodiments, compositions containing Compound 1 are administered fortherapeutic applications (e.g., administered to a subject diagnosed witha hematological malignancy). In some embodiments, compositionscontaining Compound 1 are administered for therapeutic applications(e.g., administered to a subject susceptible to or otherwise at risk ofdeveloping a hematological malignancy). In some embodiments,compositions containing Compound 1 are administered to a patient who isin remission as a maintenance therapy.

Amounts of Compound 1 will depend on the use (e.g., therapeutic,prophylactic, or maintenance). Amounts of Compound 1 will depend onseverity and course of the disease or condition, previous therapy, thepatient's health status, weight, and response to the drugs, and thejudgment of the treating physician. It is considered well within theskill of the art for one to determine such therapeutically effectiveamounts by routine experimentation (including, but not limited to, adose escalation clinical trial). In some embodiments, the amount ofCompound 1 is from 300 mg/day up to, and including, 1000 mg/day. In someembodiments, the amount of Compound 1 is from 420 mg/day up to, andincluding, 840 mg/day. In some embodiments, the amount of Compound 1 isfrom 400 mg/day up to, and including, 860 mg/day. In some embodiments,the amount of Compound 1 is about 360 mg/day. In some embodiments, theamount of Compound 1 is about 420 mg/day. In some embodiments, theamount of Compound 1 is about 560 mg/day. In some embodiments, theamount of Compound 1 is about 840 mg/day. In some embodiments, theamount of Compound 1 is from 2 mg/kg/day up to, and including, 13mg/kg/day. In some embodiments, the amount of Compound 1 is from 2.5mg/kg/day up to, and including, 8 mg/kg/day. In some embodiments, theamount of Compound 1 is from 2.5 mg/kg/day up to, and including, 6mg/kg/day. In some embodiments, the amount of Compound 1 is from 2.5mg/kg/day up to, and including, 4 mg/kg/day. In some embodiments, theamount of Compound 1 is about 2.5 mg/kg/day. In some embodiments, theamount of Compound 1 is about 8 mg/kg/day.

In some embodiments, pharmaceutical compositions described hereininclude about 140 mg of Compound 1. In some embodiments, a capsuleformulation is prepared that includes about 140 mg of Compound 1. Insome embodiments, 2, 3, 4, or 5 of the capsule formulations areadministered daily. In some embodiments, 3 or 4 of the capsules areadministered daily. In some embodiments, 3 of the 140 mg capsules areadministered once daily. In some embodiments, 4 of the 140 mg capsulesare administered once daily. In some embodiments, the capsules areadministered once daily. In other embodiments, the capsules areadministered multiple times a day.

In some embodiments, Compound 1 is administered daily. In someembodiments, Compound 1 is administered every other day.

In some embodiments, Compound 1 is administered once per day. In someembodiments, Compound 1 is administered twice per day. In someembodiments, Compound 1 is administered three times per day. In someembodiments, Compound 1 is administered four times per day.

In some embodiments, Compound 1 is administered until diseaseprogression, unacceptable toxicity, or individual choice. In someembodiments, Compound 1 is administered daily until disease progression,unacceptable toxicity, or individual choice. In some embodiments,Compound 1 is administered every other day until disease progression,unacceptable toxicity, or individual choice.

In the case wherein the patient's status does improve, upon the doctor'sdiscretion the administration of the compounds may be givencontinuously; alternatively, the dose of drug being administered may betemporarily reduced or temporarily suspended for a certain length oftime (i.e., a “drug holiday”). The length of the drug holiday can varybetween 2 days and 1 year, including by way of example only, 2 days, 3days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days,180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or365 days. The dose reduction during a drug holiday may be from 10%-100%,including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, the dosage or thefrequency of administration, or both, can be reduced, as a function ofthe symptoms, to a level at which the improved disease, disorder orcondition is retained. Patients can, however, require intermittenttreatment on a long-term basis upon any recurrence of symptoms.

The amount of a given agent that will correspond to such an amount willvary depending upon factors such as the particular compound, theseverity of the disease, the identity (e.g., weight) of the subject orhost in need of treatment, but can nevertheless be routinely determinedin a manner known in the art according to the particular circumstancessurrounding the case, including, e.g., the specific agent beingadministered, the route of administration, and the subject or host beingtreated. In general, however, doses employed for adult human treatmentwill typically be in the range of 0.02-5000 mg per day, or from about1-1500 mg per day. The desired dose may conveniently be presented in asingle dose or as divided doses administered simultaneously (or over ashort period of time) or at appropriate intervals, for example as two,three, four or more sub-doses per day.

The pharmaceutical composition described herein may be in unit dosageforms suitable for single administration of precise dosages. In unitdosage form, the formulation is divided into unit doses containingappropriate quantities of one or more compound. The unit dosage may bein the form of a package containing discrete quantities of theformulation. Non-limiting examples are packaged tablets or capsules, andpowders in vials or ampoules. Aqueous suspension compositions can bepackaged in single-dose non-reclosable containers. Alternatively,multiple-dose reclosable containers can be used, in which case it istypical to include a preservative in the composition. By way of exampleonly, formulations for parenteral injection may be presented in unitdosage form, which include, but are not limited to ampoules, or inmulti-dose containers, with an added preservative. In some embodiments,each unit dosage form comprises 140 mg of Compound 1. In someembodiments, an individual is administered 1 unit dosage form per day.In some embodiments, an individual is administered 2 unit dosage formsper day. In some embodiments, an individual is administered 3 unitdosage forms per day. In some embodiments, an individual is administered4 unit dosage forms per day.

The foregoing ranges are merely suggestive, as the number of variablesin regard to an individual treatment regime is large, and considerableexcursions from these recommended values are not uncommon. Such dosagesmay be altered depending on a number of variables, not limited to theactivity of the compound used, the disease or condition to be treated,the mode of administration, the requirements of the individual subject,the severity of the disease or condition being treated, and the judgmentof the practitioner.

Toxicity and therapeutic efficacy of such therapeutic regimens can bedetermined by standard pharmaceutical procedures in cell cultures orexperimental animals, including, but not limited to, the determinationof the LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (thedose therapeutically effective in 50% of the population). The dose ratiobetween the toxic and therapeutic effects is the therapeutic index andit can be expressed as the ratio between LD₅₀ and ED₅₀. Compoundsexhibiting high therapeutic indices are preferred. The data obtainedfrom cell culture assays and animal studies can be used in formulating arange of dosage for use in human. The dosage of such compounds liespreferably within a range of circulating concentrations that include theED₅₀ with minimal toxicity. The dosage may vary within this rangedepending upon the dosage form employed and the route of administrationutilized.

Combination Therapy

In certain instances, it is appropriate to administer Compound 1 incombination with another therapeutic agent.

In one embodiment, the compositions and methods described herein arealso used in conjunction with other therapeutic reagents that areselected for their particular usefulness against the condition that isbeing treated. In general, the compositions described herein and, inembodiments where combinational therapy is employed, other agents do nothave to be administered in the same pharmaceutical composition, and are,because of different physical and chemical characteristics, administeredby different routes. In one embodiment, the initial administration ismade according to established protocols, and then, based upon theobserved effects, the dosage, modes of administration and times ofadministration, further modified.

In various embodiments, the compounds are administered concurrently(e.g., simultaneously, essentially simultaneously or within the sametreatment protocol) or sequentially, depending upon the nature of thedisease, the condition of the patient, and the actual choice ofcompounds used. In certain embodiments, the determination of the orderof administration, and the number of repetitions of administration ofeach therapeutic agent during a treatment protocol, is based uponevaluation of the disease being treated and the condition of thepatient.

For combination therapies described herein, dosages of theco-administered compounds vary depending on the type of co-drugemployed, on the specific drug employed, on the disease or conditionbeing treated and so forth.

The individual compounds of such combinations are administered eithersequentially or simultaneously in separate or combined pharmaceuticalformulations. In one embodiment, the individual compounds will beadministered simultaneously in a combined pharmaceutical formulation.Appropriate doses of known therapeutic agents will be appreciated bythose skilled in the art.

The combinations referred to herein are conveniently presented for usein the form of a pharmaceutical compositions together with apharmaceutically acceptable diluent(s) or carrier(s).

Disclosed herein, in certain embodiments, is a method for treating acancer in an individual in need thereof, comprising: administering tothe individual an amount of Compound 1. In some embodiments, the methodfurther comprises administering a second cancer treatment regimen.

In some embodiments, administering a Btk inhibitor before a secondcancer treatment regimen reduces immune-mediated reactions to the secondcancer treatment regimen. In some embodiments, administering Compound 1before ofatumumab reduces immune-mediated reactions to ofatumumab.

In some embodiments, the second cancer treatment regimen comprises achemotherapeutic agent, a steroid, an immunotherapeutic agent, atargeted therapy, or a combination thereof. In some embodiments, thesecond cancer treatment regimen comprises a B cell receptor pathwayinhibitor. In some embodiments, the B cell receptor pathway inhibitor isa CD79A inhibitor, a CD79B inhibitor, a CD19 inhibitor, a Lyn inhibitor,a Syk inhibitor, a PI3K inhibitor, a Blnk inhibitor, a PLCγ inhibitor, aPKCβ inhibitor, or a combination thereof. In some embodiments, thesecond cancer treatment regimen comprises an antibody, B cell receptorsignaling inhibitor, a PI3K inhibitor, an IAP inhibitor, an mTORinhibitor, a radioimmunotherapeutic, a DNA damaging agent, a proteosomeinhibitor, a Cyp3A4 inhibitor, a histone deacetylase inhibitor, aprotein kinase inhibitor, a hedgehog inhibitor, an Hsp90 inhibitor, atelomerase inhibitor, a Jak1/2 inhibitor, a protease inhibitor, a PKCinhibitor, a PARP inhibitor, or a combination thereof.

In some embodiments, the second cancer treatment regimen compriseschlorambucil, ifosphamide, doxorubicin, mesalazine, thalidomide,lenalidomide, temsirolimus, everolimus, fludarabine, fostamatinib,paclitaxel, docetaxel, ofatumumab, rituximab, dexamethasone, prednisone,CAL-101, ibritumomab, tositumomab, bortezomib, pentostatin, endostatin,or a combination thereof.

In some embodiments, the second cancer treatment regimen comprisescyclophosphamide, hydroxydaunorubicin, vincristine, and prednisone, andoptionally, rituximab.

In some embodiments, the second cancer treatment regimen comprisesbendamustin, and rituximab.

In some embodiments, the second cancer treatment regimen comprisesfludarabine, cyclophosphamide, and rituximab.

In some embodiments, the second cancer treatment regimen comprisescyclophosphamide, vincristine, and prednisone, and optionally,rituximab.

In some embodiments, the second cancer treatment regimen comprisesetoposide, doxorubicin, vinristine, cyclophosphamide, prednisolone, andoptionally, rituximab.

In some embodiments, the second cancer treatment regimen comprisesdexamethasone and lenalidomide.

In some embodiments, the second cancer treatment comprises a proteasomeinhibitor. In some embodiments, the second treatment comprisesbortezomib. In some embodiments, the second cancer treatment comprisesan epoxyketone. In some embodiments, the second cancer treatmentcomprises epoxomicin. In some embodiments, the second cancer treatmentcomprises a tetrapeptide epoxyketone In some embodiments, the secondcancer treatment comprises carfilzomib. In some embodiments, the secondcancer treatment comprises disulfram, epigallocatechin-3-gallate,salinosporamide A, ONX 0912m CEP-18770, MLN9708, or MG132.

In some embodiments, the second cancer treatment comprises a Cyp3A4inhibitor. In some embodiments, the second cancer treatment comprisesindinavir, nelfinavir, ritonavir, clarithromycin, itraconazole,ketoconazole, nefazodone. In some embodiments, the second cancertreatment comprises ketoconazole.

In some embodiments, the second cancer treatment comprises a JanusKinase (JAK) inhibitor. In some embodiments, the second treatmentcomprises Lestaurtinib, Tofacitinib, Ruxolitinib, CYT387, Baricitinib orPacritinib.

In some embodiments, the second cancer treatment comprises a histonedeacetylase inhibitor (HDAC inhibitor, HDI). In some embodiments, thesecond cancer treatment comprises a hydroxamic acid (or hydroxamate),such as trichostatin A, vorinostat (SAHA), belinostat (PXD101), LAQ824,and panobinostat (LBH589), a cyclic tetrapeptide, such as trapoxin B, adepsipeptide, a benzamide, such as entinostat (MS-275), CI994, andmocetinostat (MGCD0103), an electrophilic ketone, or an aliphatic acidcompound, such as phenylbutyrate and valproic acid,

Additional cancer treatment regimens include Nitrogen Mustards such asfor example, bendamustine, chlorambucil, chlormethine, cyclophosphamide,ifosfamide, melphalan, prednimustine, trofosfamide; Alkyl Sulfonateslike busulfan, mannosulfan, treosulfan; Ethylene Imines like carboquone,thiotepa, triaziquone; Nitrosoureas like carmustine, fotemustine,lomustine, nimustine, ranimustine, semustine, streptozocin; Epoxidessuch as for example, etoglucid; Other Alkylating Agents such as forexample dacarbazine, mitobronitol, pipobroman, temozolomide; Folic AcidAnalogues such as for example methotrexate, permetrexed, pralatrexate,raltitrexed; Purine Analogs such as for example cladribine, clofarabine,fludarabine, mercaptopurine, nelarabine, tioguanine; Pyrimidine Analogssuch as for example azacitidine, capecitabine, carmofur, cytarabine,decitabine, fluorouracil, gemcitabine, tegafur; Vinca Alkaloids such asfor example vinblastine, vincristine, vindesine, vinflunine,vinorelbine; Podophyllotoxin Derivatives such as for example etoposide,teniposide; Colchicine derivatives such as for example demecolcine;Taxanes such as for example docetaxel, paclitaxel, paclitaxelpoliglumex; Other Plant Alkaloids and Natural Products such as forexample trabectedin; Actinomycines such as for example dactinomycin;Antracyclines such as for example aclarubicin, daunorubicin,doxorubicin, epirubicin, idarubicin, mitoxantrone, pirarubicin,valrubicin, zorubincin; Other Cytotoxic Antibiotics such as for examplebleomycin, ixabepilone, mitomycin, plicamycin; Platinum Compounds suchas for example carboplatin, cisplatin, oxaliplatin, satraplatin;Methylhydrazines such as for example procarbazine; Sensitizers such asfor example aminolevulinic acid, efaproxiral, methyl aminolevulinate,porfimer sodium, temoporfin; Protein Kinase Inhibitors such as forexample dasatinib, erlotinib, everolimus, gefitinib, imatinib,lapatinib, nilotinib, pazonanib, sorafenib, sunitinib, temsirolimus;Other Antineoplastic Agents such as for example alitretinoin,altretamine, amzacrine, anagrelide, arsenic trioxide, asparaginase,bexarotene, bortezomib, celecoxib, denileukin diftitox, estramustine,hydroxycarbamide, irinotecan, lonidamine, masoprocol, miltefosein,mitoguazone, mitotane, oblimersen, pegaspargase, pentostatin,romidepsin, sitimagene ceradenovec, tiazofurine, topotecan, tretinoin,vorinostat; Estrogens such as for example diethylstilbenol,ethinylestradiol, fosfestrol, polyestradiol phosphate; Progestogens suchas for example gestonorone, medroxyprogesterone, megestrol; GonadotropinReleasing Hormone Analogs such as for example buserelin, goserelin,leuprorelin, triptorelin; Anti-Estrogens such as for examplefulvestrant, tamoxifen, toremifene; Anti-Androgens such as for examplebicalutamide, flutamide, nilutamide, Enzyme Inhibitors,aminoglutethimide, anastrozole, exemestane, formestane, letrozole,vorozole; Other Hormone Antagonists such as for example abarelix,degarelix; Immunostimulants such as for example histaminedihydrochloride, mifamurtide, pidotimod, plerixafor, roquinimex,thymopentin; Immunosuppressants such as for example everolimus,gusperimus, leflunomide, mycophenolic acid, sirolimus; CalcineurinInhibitors such as for example ciclosporin, tacrolimus; OtherImmunosuppressants such as for example azathioprine, lenalidomide,methotrexate, thalidomide; and Radiopharmaceuticals such as for example,iobenguane.

Additional cancer treatment regimens include interferons, interleukins,Tumor Necrosis Factors, Growth Factors, or the like.

Additional cancer treatment regimens include Immunostimulants such asfor example ancestim, filgrastim, lenograstim, molgramostim,pegfilgrastim, sargramostim; Interferons such as for example interferonalfa natural, interferon alfa-2a, interferon alfa-2b, interferonalfacon-1, interferon alfa-n1, interferon beta natural, interferonbeta-1a, interferon beta-b, interferon gamma, peginterferon alfa-2a,peginterferon alfa-2b; Interleukins such as for example aldesleukin,oprelvekin; Other Immunostimulants such as for example BCG vaccine,glatiramer acetate, histamine dihydrochloride, immunocyanin, lentinan,melanoma vaccine, mifamurtide, pegademase, pidotimod, plerixafor, polyI:C, poly ICLC, roquinimex, tasonermin, thymopentin; Immunosuppressantssuch as for example abatacept, abetimus, alefacept, antilymphocyteimmunoglobulin (horse), antithymocyte immunoglobulin (rabbit),eculizumab, efalizumab, everolimus, gusperimus, leflunomide,muromab-CD3, mycophenolic acid, natalizumab, sirolimus; TNF alphaInhibitors such as for example adalimumab, afelimomab, certolizumabpegol, etanercept, golimumab, infliximab; Interleukin Inhibitors such asfor example anakinra, basiliximab, canakinumab, daclizumab, mepolizumab,rilonacept, tocilizumab, ustekinumab; Calcineurin Inhibitors such as forexample ciclosporin, tacrolimus; Other Immunosuppressants such as forexample azathioprine, lenalidomide, methotrexate, thalidomide.

Additional cancer treatment regimens include Adalimumab, Alemtuzumab,Basiliximab, Bevacizumab, Cetuximab, Certolizumab pegol, Daclizumab,Eculizumab, Efalizumab, Gemtuzumab, Ibritumomab tiuxetan, Infliximab,Muromonab-CD3, Natalizumab, Panitumumab, Ranibizumab, Rituximab,Tositumomab, Trastuzumab, or the like, or a combination thereof.

Additional cancer treatment regimens include Monoclonal Antibodies suchas for example alemtuzumab, bevacizumab, catumaxomab, cetuximab,edrecolomab, gemtuzumab, ofatumumab, panitumumab, rituximab,trastuzumab, Immunosuppressants, eculizumab, efalizumab, muromab-CD3,natalizumab; TNF alpha Inhibitors such as for example adalimumab,afelimomab, certolizumab pegol, golimumab, infliximab, InterleukinInhibitors, basiliximab, canakinumab, daclizumab, mepolizumab,tocilizumab, ustekinumab, Radiopharmaceuticals, ibritumomab tiuxetan,tositumomab; Others Monoclonal Antibodies such as for exampleabagovomab, adecatumumab, alemtuzumab, anti-CD30 monoclonal antibodyXmab2513, anti-MET monoclonal antibody MetMab, apolizumab, apomab,arcitumomab, basiliximab, bispecific antibody 2B1, blinatumomab,brentuximab vedotin, capromab pendetide, cixutumumab, claudiximab,conatumumab, dacetuzumab, denosumab, eculizumab, epratuzumab,epratuzumab, ertumaxomab, etaracizumab, figitumumab, fresolimumab,galiximab, ganitumab, gemtuzumab ozogamicin, glembatumumab, ibritumomab,inotuzumab ozogamicin, ipilimumab, lexatumumab, lintuzumab, lintuzumab,lucatumumab, mapatumumab, matuzumab, milatuzumab, monoclonal antibodyCC49, necitumumab, nimotuzumab, ofatumumab, oregovomab, pertuzumab,ramacurimab, ranibizumab, siplizumab, sonepcizumab, tanezumab,tositumomab, trastuzumab, tremelimumab, tucotuzumab celmoleukin,veltuzumab, visilizumab, volociximab, zalutumumab.

Additional cancer treatment regimens include agents that affect thetumor micro-environment such as cellular signaling network (e.g.phosphatidylinositol 3-kinase (PI3K) signaling pathway, signaling fromthe B-cell receptor and the IgE receptor). In some embodiments, thesecond agent is a PI3K signaling inhibitor or a syc kinase inhibitor. Inone embodiment, the syk inhibitor is R788. In another embodiment is aPKCγ inhibitor such as by way of example only, enzastaurin.

Examples of agents that affect the tumor micro-environment include PI3Ksignaling inhibitor, syc kinase inhibitor, Protein Kinase Inhibitorssuch as for example dasatinib, erlotinib, everolimus, gefitinib,imatinib, lapatinib, nilotinib, pazonanib, sorafenib, sunitinib,temsirolimus; Other Angiogenesis Inhibitors such as for example GT-111,JI-101, R1530; Other Kinase Inhibitors such as for example AC220, AC480,ACE-041, AMG 900, AP24534, Arry-614, AT7519, AT9283, AV-951, axitinib,AZD1152, AZD7762, AZD8055, AZD8931, bafetinib, BAY 73-4506, BGJ398,BGT226, BI 811283, BI6727, BIBF 1120, BIBW 2992, BMS-690154, BMS-777607,BMS-863233, BSK-461364, CAL-101, CEP-11981, CYC116, DCC-2036,dinaciclib, dovitinib lactate, E7050, EMD 1214063, ENMD-2076,fostamatinib disodium, GSK2256098, GSK690693, INCB18424, INNO-406,JNJ-26483327, JX-594, KX2-391, linifanib, LY2603618, MGCD265, MK-0457,MK1496, MLN8054, MLN8237, MP470, NMS-1116354, NMS-1286937, ON 01919.Na,OSI-027, OSI-930, Btk inhibitor, PF-00562271, PF-02341066, PF-03814735,PF-04217903, PF-04554878, PF-04691502, PF-3758309, PHA-739358, PLC3397,progenipoietin, R547, R763, ramucirumab, regorafenib, R05185426,SAR103168, SCH 727965, SGI-1176, SGX523, SNS-314, TAK-593, TAK-901,TKI258, TLN-232, TTP607, XL147, XL228, XL281RO5126766, XL418, XL765.

Further examples of anti-cancer agents for use in combination with a Btkinhibitor compound include inhibitors of mitogen-activated proteinkinase signaling, e.g., U0126, PD98059, PD184352, PD0325901,ARRY-142886, SB239063, SP600125, BAY 43-9006, wortmannin, or LY294002;Syk inhibitors; mTOR inhibitors; and antibodies (e.g., rituxan).

Other anti-cancer agents that can be employed in combination with a Btkinhibitor compound include Adriamycin, Dactinomycin, Bleomycin,Vinblastine, Cisplatin, acivicin; aclarubicin; acodazole hydrochloride;acronine; adozelesin; aldesleukin; altretamine; ambomycin; ametantroneacetate; aminoglutethimide; amsacrine; anastrozole; anthramycin;asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat;benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate;bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefingol;chlorambucil; cirolemycin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride;decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate;diaziquone; doxorubicin; doxorubicin hydrochloride; droloxifene;droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate;eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate;epipropidine; epirubicin hydrochloride; erbulozole; esorubicinhydrochloride; estramustine; estramustine phosphate sodium; etanidazole;etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride;fazarabine; fenretinide; floxuridine; fludarabine phosphate;fluorouracil; flurocitabine; fosquidone; fostriecin sodium; gemcitabine;gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride;ifosfamide; iimofosine; interleukin II (including recombinantinterleukin II, or rlL2), interferon alfa-2a; interferon alfa-2b;interferon alfa-n1; interferon alfa-n3; interferon beta-1 a; interferongamma-1 b; iproplatin; irinotecan hydrochloride; lanreotide acetate;letrozole; leuprolide acetate; liarozole hydrochloride; lometrexolsodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine;mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate;melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium;metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin;mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride;mycophenolic acid; nocodazoie; nogalamycin; ormaplatin; oxisuran;pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride;semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantronehydrochloride; temoporfin; teniposide; teroxirone; testolactone;thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifenecitrate; trestolone acetate; triciribine phosphate; trimetrexate;trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracilmustard; uredepa; vapreotide; verteporfin; vinblastine sulfate;vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate;vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate;vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin;zinostatin; zorubicin hydrochloride.

Other anti-cancer agents that can be employed in combination with a Btkinhibitor compound include: 20-epi-1, 25 dihydroxyvitamin D3;5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol;adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine;amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine;anagrelide; anastrozole; andrographolide; angiogenesis inhibitors;antagonist D; antagonist G; antarelix; anti-dorsalizing morphogeneticprotein-1; antiandrogen, prostatic carcinoma; antiestrogen;antineoplaston; antisense oligonucleotides; aphidicolin glycinate;apoptosis gene modulators; apoptosis regulators; apurinic acid;ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane;atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron;azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat;BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactamderivatives; beta-alethine; betaclamycin B; betulinic acid; bFGFinhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide;bistratene A; bizelesin; breflate; bropirimine; budotitane; buthioninesulfoximine; calcipotriol; calphostin C; camptothecin derivatives;canarypox IL-2; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorlns; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron;doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen;ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur;epirubicin; epristeride; estramustine analogue; estrogen agonists;estrogen antagonists; etanidazole; etoposide phosphate; exemestane;fadrozole; fazarabine; fenretinide; filgrastim; finasteride;flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-such asfor example growth factor-1 receptor inhibitor; interferon agonists;interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-;iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin;pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine;pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin;pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylerie conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen-binding protein; sizofiran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer.

Yet other anticancer agents that can be employed in combination with aBtk inhibitor compound include alkylating agents, antimetabolites,natural products, or hormones, e.g., nitrogen mustards (e.g.,mechloroethamine, cyclophosphamide, chlorambucil, etc.), alkylsulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomusitne,etc.), or triazenes (decarbazine, etc.). Examples of antimetabolitesinclude but are not limited to folic acid analog (e.g., methotrexate),or pyrimidine analogs (e.g., Cytarabine), purine analogs (e.g.,mercaptopurine, thioguanine, pentostatin).

Examples of alkylating agents that can be employed in combination a Btkinhibitor compound include, but are not limited to, nitrogen mustards(e.g., mechloroethamine, cyclophosphamide, chlorambucil, meiphalan,etc.), ethylenimine and methylmelamines (e.g., hexamethlymelamine,thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g.,carmustine, lomusitne, semustine, streptozocin, etc.), or triazenes(decarbazine, etc.). Examples of antimetabolites include, but are notlimited to folic acid analog (e.g., methotrexate), or pyrimidine analogs(e.g., fluorouracil, floxouridine, Cytarabine), purine analogs (e.g.,mercaptopurine, thioguanine, pentostatin.

Examples of anti-cancer agents which act by arresting cells in the G2-Mphases due to stabilized microtubules and which can be used incombination with a Btk inhibitor compound include without limitation thefollowing marketed drugs and drugs in development: Erbulozole (alsoknown as R-55104), Dolastatin 10 (also known as DLS-10 and NSC-376128),Mivobulin isethionate (also known as CI-980), Vincristine, NSC-639829,Discodermolide (also known as NVP-XX-A-296), ABT-751 (Abbott, also knownas E-7010), Altorhyrtins (such as Altorhyrtin A and Altorhyrtin C),Spongistatins (such as Spongistatin 1, Spongistatin 2, Spongistatin 3,Spongistatin 4, Spongistatin 5, Spongistatin 6, Spongistatin 7,Spongistatin 8, and Spongistatin 9), Cemadotin hydrochloride (also knownas LU-103793 and NSC-D-669356), Epothilones (such as Epothilone A,Epothilone B, Epothilone C (also known as desoxyepothilone A or dEpoA),Epothilone D (also referred to as KOS-862, dEpoB, and desoxyepothiloneB), Epothilone E, Epothilone F, Epothilone B N-oxide, Epothilone AN-oxide, 16-aza-epothilone B, 21-aminoepothilone B (also known asBMS-310705), 21-hydroxyepothilone D (also known as Desoxyepothilone Fand dEpoF), 26-fluoroepothilone), Auristatin PE (also known asNSC-654663), Soblidotin (also known as TZT-1027), LS-4559-P (Pharmacia,also known as LS-4577), LS-4578 (Pharmacia, also known as LS-477-P),LS-4477 (Pharmacia), LS-4559 (Pharmacia), RPR-112378 (Aventis),Vincristine sulfate, DZ-3358 (Daiichi), FR-182877 (Fujisawa, also knownas WS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2 (Hungarian Academyof Sciences), BSF-223651 (BASF, also known as ILX-651 and LU-223651),SAH-49960 (Lilly/Novartis), SDZ-268970 (Lilly/Novartis), AM-97(Armad/Kyowa Hakko), AM-132 (Armad), AM-138 (Armad/Kyowa Hakko),IDN-5005 (Indena), Cryptophycin 52 (also known as LY-355703), AC-7739(Ajinomoto, also known as AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto,also known as AVE-8062, AVE-8062A, CS-39-L-Ser.HCl, and RPR-258062A),Vitilevuamide, Tubulysin A, Canadensol, Centaureidin (also known asNSC-106969), T-138067 (Tularik, also known as T-67, TL-138067 andTI-138067), COBRA-1 (Parker Hughes Institute, also known as DDE-261 andWHI-261), H10 (Kansas State University), H16 (Kansas State University),Oncocidin A1 (also known as BTO-956 and DIME), DDE-313 (Parker HughesInstitute), Fijianolide B, Laulimalide, SPA-2 (Parker Hughes Institute),SPA-1 (Parker Hughes Institute, also known as SPIKET-P), 3-IAABU(Cytoskeleton/Mt. Sinai School of Medicine, also known as MF-569),Narcosine (also known as NSC-5366), Nascapine, D-24851 (Asta Medica),A-105972 (Abbott), Hemiasterlin, 3-BAABU (Cytoskeleton/Mt. Sinai Schoolof Medicine, also known as MF-191), TMPN (Arizona State University),Vanadocene acetylacetonate, T-138026 (Tularik), Monsatrol, lnanocine(also known as NSC-698666), 3-1AABE (Cytoskeleton/Mt. Sinai School ofMedicine), A-204197 (Abbott), T-607 (Tuiarik, also known as T-900607),RPR-115781 (Aventis), Eleutherobins (such as Desmethyleleutherobin,Desaetyleleutherobin, lsoeleutherobin A, and Z-Eleutherobin),Caribaeoside, Caribaeolin, Halichondrin B, D-64131 (Asta Medica),D-68144 (Asta Medica), Diazonamide A, A-293620 (Abbott), NPI-2350(Nereus), Taccalonolide A, TUB-245 (Aventis), A-259754 (Abbott),Diozostatin, (−)-Phenylahistin (also known as NSCL-96F037), D-68838(Asta Medica), D-68836 (Asta Medica), Myoseverin B, D-43411 (Zentaris,also known as D-81862), A-289099 (Abbott), A-318315 (Abbott), HTI-286(also known as SPA-110, trifluoroacetate salt) (Wyeth), D-82317(Zentaris), D-82318 (Zentaris), SC-12983 (NCI), Resverastatin phosphatesodium, BPR-OY-007 (National Health Research Institutes), and SSR-250411(Sanofi).

Where the individual is suffering from or at risk of suffering from anautoimmune disease, an inflammatory disease, or an allergy disease,Compound 1 can be used in with one or more of the following therapeuticagents in any combination: immunosuppressants (e.g., tacrolimus,cyclosporin, rapamicin, methotrexate, cyclophosphamide, azathioprine,mercaptopurine, mycophenolate, or FTY720), glucocorticoids (e.g.,prednisone, cortisone acetate, prednisolone, methylprednisolone,dexamethasone, betamethasone, triamcinolone, beclometasone,fludrocortisone acetate, deoxycorticosterone acetate, aldosterone),non-steroidal anti-inflammatory drugs (e.g., salicylates, arylalkanoicacids, 2-arylpropionic acids, N-arylanthranilic acids, oxicams, coxibs,or sulphonanilides), Cox-2-specific inhibitors (e.g., valdecoxib,celecoxib, or rofecoxib), leflunomide, gold thioglucose, goldthiomalate, aurofin, sulfasalazine, hydroxychloroquinine, minocycline,TNF-α binding proteins (e.g., infliximab, etanercept, or adalimumab),abatacept, anakinra, interferon-β, interferon-γ, interleukin-2, allergyvaccines, antihistamines, antileukotrienes, beta-agonists, theophylline,or anticholinergics.

Kits/Articles of Manufacture

For use in the therapeutic methods of use described herein, kits andarticles of manufacture are also described herein. Such kits include acarrier, package, or container that is compartmentalized to receive oneor more containers such as vials, tubes, and the like, each of thecontainer(s) comprising one of the separate elements to be used in amethod described herein. Suitable containers include, for example,bottles, vials, syringes, and test tubes. In one embodiment, thecontainers are formed from a variety of materials such as glass orplastic.

The articles of manufacture provided herein contain packaging materials.Packaging materials for use in packaging pharmaceutical productsinclude, e.g., U.S. Pat. No. 5,323,907. Examples of pharmaceuticalpackaging materials include, but are not limited to, blister packs,bottles, tubes, bags, containers, bottles, and any packaging materialsuitable for a selected formulation and intended mode of administrationand treatment.

In some embodiments, the compounds or compositions described herein, arepresented in a package or dispenser device which may contain one or moreunit dosage forms containing the active ingredient. The compound orcomposition described herein is packaged alone, or packaged with anothercompound or another ingredient or additive. In some embodiments, thepackage contains one or more containers filled with one or more of theingredients of the pharmaceutical compositions. In some embodiments, thepackage comprises metal or plastic foil, such as a blister pack. In someembodiments, the package or dispenser device is accompanied byinstructions for administration, such as instructions for administeringthe compounds or compositions for treating a neoplastic disease. In someembodiments, the package or dispenser is accompanied with a noticeassociated with the container in form prescribed by a governmentalagency regulating the manufacture, use, or sale of pharmaceuticals,which notice is reflective of approval by the agency of the form of thedrug for human or veterinary administration. In some embodiments, suchnotice, for example, is the labeling approved by the U.S. Food and DrugAdministration for prescription drugs, or the approved product insert.In some embodiments, compositions include a compound described hereinformulated in a compatible pharmaceutical carrier are prepared, placedin an appropriate container, and labeled for treatment of an indicatedcondition.

For example, the container(s) include Compound 1, optionally in acomposition or in combination with another agent as disclosed herein.Such kits optionally include an identifying description or label orinstructions relating to its use in the methods described herein.

A kit typically includes labels listing contents and/or instructions foruse, and package inserts with instructions for use. A set ofinstructions will also typically be included.

In one embodiment, a label is on or associated with the container. Inone embodiment, a label is on a container when letters, numbers or othercharacters forming the label are attached, molded or etched into thecontainer itself; a label is associated with a container when it ispresent within a receptacle or carrier that also holds the container,e.g., as a package insert. In one embodiment, a label is used toindicate that the contents are to be used for a specific therapeuticapplication. The label also indicates directions for use of thecontents, such as in the methods described herein.

In certain embodiments, the pharmaceutical compositions are presented ina pack or dispenser device which contains one or more unit dosage formscontaining a compound provided herein. The pack, for example, containsmetal or plastic foil, such as a blister pack. In one embodiment, thepack or dispenser device is accompanied by instructions foradministration. In one embodiment, the pack or dispenser is alsoaccompanied with a notice associated with the container in formprescribed by a governmental agency regulating the manufacture, use, orsale of pharmaceuticals, which notice is reflective of approval by theagency of the form of the drug for human or veterinary administration.Such notice, for example, is the labeling approved by the U.S. Food andDrug Administration for prescription drugs, or the approved productinsert. In one embodiment, compositions containing a compound providedherein formulated in a compatible pharmaceutical carrier are alsoprepared, placed in an appropriate container, and labeled for treatmentof an indicated condition.

EXAMPLES

The following ingredients, formulations, processes and procedures forpracticing the methods disclosed herein correspond to that describedabove.

Example 1: Preparation of Crystalline Forms of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one(Compound 1)

Form A—Route 1:

Amorphous Compound 1 (ca. 15 mg) was measured into a vial. Ten volumes(150 μl) of solvent [methyl tert-butyl ether (MTBE), diisopropyl ether(DIPE), ethyl acetate, isopropyl acetate, isopropyl alcohol, methylisobutyl ketone (MIBK), methyl ethyl ketone (MEK), acetone, methanol,nitromethane, 10% aqueous acetone, or 10% aqueous isopropyl alcohol]were added to the vial. The vial was sealed and placed in a shaker at50° C. for one hour. If a slurry was obtained, an additional thirtyvolumes (total of 600 μl) of solvent was added, then the slurry wasreturned to 50° C. for another hour. If the sample remained as a slurryat this point, no further solvent was added. The solution/slurry wasstirred at 50° C. for one hour, then cooled to 0° C. at 0.1° C./min,then held at 0° C. overnight. If a slurry was obtained, the solids werefiltered under vacuum to provide Compound 1, Form A; the solution wasreturned to ambient temperature for slow evaporation through a pin-holeto furnish Compound 1, Form A.

Form A—Route 2:

Amorphous Compound 1 (20 mg) was added to a vial, followed by solvent[heptanes (10 volumes), dioxane (1 volume), toluene (10 volumes), MTBE(10 volumes), DIPE (10 volumes), anisole (1 volume), ethyl acetate (10volumes), isopropyl acetate (10 volumes), tetrahydrofuran (1 volume),DCM (1 volume), MIBK (10 volumes), MEK (10 volumes), acetone (10volumes), methanol (10 volumes), ethanol (10 volumes), acetonitrile (10volumes), nitromethane (1 volume), water (10 volumes), or 10% aqueousisopropyl alcohol (1 volume)]. The sealed vial was placed in amaturation chamber (cycling between 50° C. and ambient for four hourseach) for five days before the solids were filtered under vacuum toprovide Compound 1, Form A.

In some embodiments, amorphous Compound 1 is prepared by dissolvingCompound 1, Form A (ca. 500 mg) in 10 ml dichloromethane (DCM). Thesolvent was removed by rotary evaporation, occurring rapidly enough toprevent crystallization to provide amorphous Compound 1.

Form A—Route 3:

In a clean round bottom flask, 12.0 grams of Compound 1 were dissolvedin 120 ml of methanol by heating to 45° C. with magnetic stirring. Tothe warm solution of dissolved Compound 1 was added 72 ml of water over45 min, maintaining the internal temperature at 45° C. The solutionslowly became a slurry and was stirred for 3 hours at the elevatedtemperature. A sample of the slurry was drawn, filtered and dried. Theslurry was allowed to cool to room temperature and stirred for at least16 hours. Another sample of the slurry was drawn, filtered and dried.The solids were filtered, washed with 50 ml of a 3:2 mixture ofmethanol: water, and dried on the filter for 40 hours. Yield 9.6 gramsof Form A (melting points: 1^(st) sample ˜152° C., 2^(nd) sample ˜154°C., main lot ˜154° C.).

Form A was also attained in an analogous manner using aqueous acetone,ethanol, and n-propanol.

Form B—Route 1:

Compound 1, Form A (ca. 100 mg) was weighed into a vessel and dissolvedin methanol (2 ml). The solution was heated to 50° C. to ensure fulldissolution, then cooled to 5° C. Water was added to the solution at 5°C. (200 μl at a time until sample became cloudy, totaling 1000 μl).Seeds of Compound 1, Form C were added immediately upon appearance ofcloudiness. The slurry was stirred at 5° C. for one day. An aliquot wasremoved by pipette for analysis by XRPD, keeping the bulk of the samplein the same conditions. The XRPD analysis highlighted the lowcrystallinity of the material, so the sample was held at 5° C. for anadditional three days. After this time, re-analysis of an aliquot of thesample showed the material to have converted to Compound 1, Form B. Thesample was isolated by filtration under vacuum to provide Compound 1,Form B.

Form B—Route 2:

Compound 1, Form A (ca. 500 mg) was weighed into a vessel and dissolvedin methanol (4 ml) at 50° C. The solution was cooled to 25° C.,remaining in solution. Water was added (500 μl of water added at a time,2 ml in total) until the solution became cloudy. The slurry was stirredfor ten minutes. An aliquot was removed by pipette to assess thematerial by XRPD while the sample stirred at 25° C. for an hour;however, the material was of very low crystallinity. After the hourstirring at 25° C., the sample was placed at 5° C. for three days. Afterthis time, another aliquot was removed by pipette for XRPD analysis. Theremainder of the slurry was filtered under vacuum and dried at 25° C.under vacuum overnight to provide Compound 1, Form B.

Form C:

In a clean round bottom flask, 2.0 grams of Compound 1 were suspended in25 ml of methanol and heated to 50° C. The warm solution of dissolvedCompound 1 was filtered into a clean round bottom flask. The polishedsolution was allowed to cool to room temperature with magnetic stirring.The solution slowly became a slurry and was stirred for over 14 hours.The solids were filtered, washed with 5 mL of methanol, and dried on thefilter for 20 hours, then at 50° C. in a vacuum-oven for 8 hours. Yield1.4 grams of Form C (melting point=−132° C.).

Form D:

A dry mixture (ca. 5 mg of each component) was prepared using two ofForm A, Form B, or Form C of Compound 1. A slurry made from amorphousCompound 1 in MIBK was filtered to obtain a saturated solution. Tenvolumes (100 μl) of the saturated solution were added to the dry mixtureto prepare a new slurry. The slurry was stored at 5° C. for three daysbefore filtration under vacuum to provide Compound 1, Form D.

Form E:

Amorphous compound 1 (20 mg) was added to a vial, followed by seeds ofCompound 1, Form C (ca. 5 mg). Ten volumes of toluene (200 μl) wereadded to the vial to prepare a slurry. The vial was sealed and matured(cycling between 50° C. and ambient temperature for four hours each) forone day. An aliquot was removed by pipette for analysis by XRPD, TGA,and DSC; data were consistent with Compound 1, Form E. However, thiscompound was found to have converted to Compound 1, Form A afterstanding at ambient temperature overnight and drying at 40° C. in vacuofor one day.

Form F:

In a clean 20 ml scintillation vial, 200 mgs of Compound 1 and 50 mgs ofactivated charcoal were suspended in 4 ml of methanol and heated to 50°C. The resulting mixture was stirred at 50° C. for 2.5 hours. The warmsolution of dissolved Compound 1 was filtered though a syringe filterinto a new, clean 20 ml vial, removing the charcoal. The polishedsolution was allowed to cool to room temperature. Without stirring, thesolution was aged for over a week when some crystals were observed tohave formed. After 6 more weeks the bottom of the vial was covered withlarge crystals. The crystals were maintained under the supersaturatedmethanol solution for analysis.

Example 2: X-Ray Powder Diffraction (XRPD)

X-Ray powder diffraction patterns were collected on a Bruker AXS C2GADDS or Bruker AXS D8 diffractometer.

Bruker AXS C2 GADDS

X-Ray Powder Diffraction patterns were collected on a Bruker AXS C2GADDS diffractometer using Cu Ka radiation (40 kV, 40 mA), automated XYZstage, laser video microscope for auto-sample positioning and a HiStar2-dimensional area detector. X-ray optics consists of a single Göbelmultilayer mirror coupled with a pinhole collimator of 0.3 mm. A weeklyperformance check is carried out using a certified standard NIST 1976Corundum (flat plate). The beam divergence, i.e. the effective size ofthe X-ray beam on the sample, was approximately 4 mm. A θ-θ continuousscan mode was employed with a sample—detector distance of 20 cm whichgives an effective 20 range of 3.2°-29.7°. Typically the sample would beexposed to the X-ray beam for 120 seconds. The software used for datacollection was GADDS for WNT 4.1.16 and the data were analysed andpresented using Diffrac Plus EVA v11.0.0.2 or v13.0.0.2.

Ambient Conditions

Samples run under ambient conditions were prepared as flat platespecimens using powder as received without grinding. Approximately 1-2mg of the sample was lightly pressed on a glass slide to obtain a flatsurface.

Non-Ambient Conditions

Samples run under non-ambient conditions were mounted on a silicon waferwith heatconducting compound. The sample was then heated to theappropriate temperature at 10° C./min (unless otherwise stated) andsubsequently held isothermally for 1 minute before data collection wasinitiated.

Bruker AXS D8 Advance

X-Ray Powder Diffraction patterns were collected on a Bruker D8diffractometer using Cu Ka radiation (40 kV, 40 mA), θ-2θ goniometer,and divergence of V4 and receiving slits, a Ge monochromator and aLynxeye detector. The instrument is performance checked using acertified Corundum standard (NIST 1976). The software used for datacollection was Diffrac Plus XRD Commander v2.5.0 and the data wereanalysed and presented using Diffrac Plus EVA v11.0.0.2 or v13.0.0.2.Samples were run under ambient conditions as flat plate specimens usingpowder as received. The sample was gently packed into a cavity cut intopolished, zero-background (510) silicon wafer. The sample was rotated inits own plane during analysis. The details of the data collection are:

-   -   Angular range: 2 to 42°2θ    -   Step size: 0.05°2θ    -   Collection time: 0.5 s/step

XRPD on Form A

The X-Ray powder diffraction for Form A is displayed in FIG. 1.Characteristic peaks include 5.7±0.1° 2-Theta, 13.6±0.1° 2-Theta,16.1±0.1° 2-Theta, 18.9±0.1° 2-Theta, 21.3±0.1° 2-Theta, and 21.6±0.1°2-Theta.

Crystallinity was unaffected after one week storage at 40° C./75% RH orafter one week storage at 25° C./97% RH.

XRPD on Form B

The X-Ray powder diffraction for Form B is displayed in FIG. 5.Characteristic peaks include 5.2±0.1° 2-Theta, 10.2±0.1° 2-Theta,16.5±0.1° 2-Theta, 18.5±0.1° 2-Theta, and 20.8±0.1° 2-Theta.

Crystallinity was unaffected after one week storage at 40° C./75% RH orafter one week storage at 25° C./97% RH.

XRPD on Form C

The X-Ray powder diffraction for Form C is displayed in FIG. 9.Characteristic peaks include 7.0±0.1° 2-Theta, 14.0±0.1° 2-Theta,15.7±0.1° 2-Theta, 18.2±0.1° 2-Theta, 19.1±0.1° 2-Theta, 19.5±0.1°2-Theta, 20.3±0.1° 2-Theta, 22.1±0.1° 2-Theta, and 22.9±0.1° 2-Theta.

Crystallinity was unaffected after one week storage at 40° C./75% RH orafter one week storage at 25° C./97% RH.

XRPD on Form D

The X-Ray powder diffraction for Form D is displayed in FIG. 12.Characteristic peaks include 7.2±0.1° 2-Theta, 8.0±0.1° 2-Theta,9.2±0.1° 2-Theta, 14.5±0.1° 2-Theta, 18.5±0.1° 2-Theta, 19.5±0.1°2-Theta, 20.7±0.1° 2-Theta, 21.0±0.1° 2-Theta, 21.9±0.1° 2-Theta, and22.4±0.1° 2-Theta.

XRPD on Form E

The X-Ray powder diffraction for Form E is displayed in FIG. 14.Characteristic peaks include 7.8±0.1° 2-Theta, 8.8±0.1° 2-Theta,16.1±0.1° 2-Theta, 18.1±0.1° 2-Theta, 19.3±0.1° 2-Theta, 19.5±0.1°2-Theta, 20.5±0.1° 2-Theta, 21.6±0.1° 2-Theta, and 25.2±0.1° 2-Theta.

XRPD on Form F

A simulated XRPD pattern was generated for Form F. The XRPD simulatedpattern was generated from the single crystal data obtained from Example3 (cif file) using Mercury CSD v3.1 (C. F. Macrae et al. J. Appl. Cryst.(2006), 39-3, 453-457) (XRPD pattern settings: CuKα 1.54056; Start/End2/43 2θ°; PWHW (2θ°) 0.1). The data was then saved as a raw file. Togenerate the 20°/Intensity (%) peak table the raw file was treated usingDiffrac Plus EVA v. 15,0,0,0.

The X-Ray powder diffraction simulated pattern for Form F is displayedin FIG. 16.

Characteristic peaks include 6.2±0.1° 2-Theta, 10.1±0.1° 2-Theta,17.6±0.1° 2-Theta, 18.6±0.1° 2-Theta, 20.0±0.1° 2-Theta, 20.4±0.1°2-Theta, 20.7±0.1° 2-Theta, 22.4±0.1° 2-Theta, 23.0±0.1° 2-Theta,23.2±0.1° 2-Theta, 24.4±0.1° 2-Theta, 25.1±0.1° 2-Theta, 27.6±0.1°2-Theta, 29.3±0.1° 2-Theta, and 29.7±0.1° 2-Theta.

Example 3: Single Crystal X-Ray Diffraction

Single crystal X-ray diffraction data was collected and processed asfollows:

Diffractometer SuperNova, Dual, Cu at zero, Atlas

Radiation source SuperNova (Cu) X-ray Source, CuKα

Data collection method omega scans

Theta range for data collection 9.11 to 74.49°

Index ranges −11≤h≤12, −12≤k≤12, −18≤l≤18

Reflections collected 22842

Independent reflections 9278 [R(int)=0.0407]

Coverage of independent reflections 99.4%

Variation in check reflections N/A

Absorption correction Semi-empirical from equivalents

Max. and min. transmission 1.00000 and 0.73583

Structure solution technique direct

Structure solution program SHELXS-97 (Sheldrick, 1990)

Refinement technique Full-matrix least-squares on F²

Refinement program SHELXL-97 (Sheldrick, 1997)

Function minimized Σw(F_(o) ²−F_(c) ²)²

Data/restraints/parameters 9278/3/660

Goodness-of-fit on F² 1.004

Δ/σ_(max) 0.000

Final R indices

9185 data; I>2σ(I) R1=0.0414, wR2=0.1144

all data R1=0.0417, wR2=0.1149

Weighting scheme calc w=1/[σ2 (F_(o) ²)+(0.0810P)²+0.2800P]

-   -   where P=(F_(o) ²+2F_(c) ²)/3        Absolute structure parameter −0.01(13)        Extinction coefficient 0.0013(3)        Largest diff. peak and hole 0.320 and −0.285 eÅ⁻³

Form F was characterized by unit cell unit cell parameters approximatelyequal to the following at a temperature of approximately 100(2) K:

Molecular formula C₅₃H₆₀N₁₂O₇ Molecular weight 977.13 Crystal systemTriclinic Space group P1 a 9.6332(3) Å α 105.762(3)° b 9.7536(4) Å β 95.132(2)° c 15.0592(4) Å  γ 111.332(3)° V 1240.15(7) Å³ Z 1 Density1.308 Mg/m³ (calculated) Absorption 0.726 mm⁻¹ coefficient Wavelength1.54178 Å F(000) 518 T 100(2) K

Example 4: Fourier Transform—Infra-Red (FTIR)

Data were collected on a Perkin-Elmer Spectrum One fitted with auniversal Attenuated Total Reflectance (ATR) sampling accessory. Thedata were collected and analysed using Spectrum v5.0.1 software.

The infrared spectrum for Form A is displayed in FIG. 2. Characteristicpeaks observed in the infrared spectrum for Form A include peaks at:1584 cm⁻¹, 1240 cm⁻¹, 1147 cm⁻¹, 1134 cm⁻¹, 1099 cm⁻¹, 1067 cm⁻¹, 985cm⁻¹, and 953 cm⁻¹.

The infrared spectrum for Form B is displayed in FIG. 6. Characteristicpeaks observed in the infrared spectrum for Form B include peaks at:1586 cm⁻¹, 1573 cm⁻¹, 1562 cm⁻¹, 1229 cm⁻¹, 1166 cm⁻¹, 1141 cm⁻¹, 1103cm⁻¹, 1056 cm⁻¹, 1033 cm⁻¹, and 982 cm⁻¹.

Example 5: Differential Scanning Calorimetry (DSC) andThermo-Gravimetric Analysis (TGA)

DSC data were collected on a TA Instruments Q2000 equipped with a 50position autosampler. The calibration for thermal capacity was carriedout using sapphire and the calibration for energy and temperature wascarried out using certified indium. Typically 0.5-3 mg of each sample,in a pin-holed aluminium pan, was heated at 10° C./min from 25° C. to300° C. A purge of dry nitrogen at 50 ml/min was maintained over thesample, unless otherwise stated. Modulated temperature DSC was carriedout using an underlying heating rate of 2° C./min and temperaturemodulation parameters of ±0.318° C. (amplitude) every 60 seconds(period). The instrument control software was Advantage for Q Seriesv2.8.0.392 and Thermal Advantage v4.8.3 and the data were analysed usingUniversal Analysis v4.4A.

TGA data were collected on a TA Instruments Q500 TGA, equipped with a 16position autosampler. The instrument was temperature calibrated usingcertified Alumel and Nickel. Typically 3-10 mg of each sample was loadedonto a pre-tared aluminium DSC pan and heated at 10° C./min from ambienttemperature to 350° C. A nitrogen purge at 60 ml/min was maintained overthe sample, unless otherwise stated. The instrument control software wasAdvantage for Q Series v2.8.0.392 and Thermal Advantage v4.8.3 and thedata were analysed using Universal Analysis v4.4A.

Form A

DSC and TGA thermograms for Form A are displayed in FIG. 3 and FIG. 4respectively.

No weight loss was observed. Material is anhydrous.

In the DSC (rate of heating: 10° C./min or 20° C./min) an endotherm wasobserved having an onset at about 154° C. and a peak at about 157° C. Anexothermic peak was observed at 159° C.

Form B

DSC and TGA thermograms for Form B are displayed in FIG. 7 and FIG. 8respectively.

No weight loss was observed. Material is anhydrous.

In the DSC (rate of heating: 10° C./min or 20° C./min) an endotherm wasobserved having an onset at about 99-106° C. and a peak at about115-118° C.

Form C

DSC and TGA thermograms for Form C are displayed in FIG. 10 and FIG. 11respectively.

No weight loss was observed. Material is anhydrous.

In the DSC (rate of heating: 10° C./min or 20° C./min) an endotherm wasobserved having an onset at about 134-135° C. and a peak at about137-139° C.

Form D

A TGA thermogram for Form D is displayed in FIG. 13.

Total weight loss of 16.6-17.8%, equivalent to about one mole of MIBK,was observed by TGA either as 1 step or as 2 steps.

Form E

DSC and TGA thermograms for Form E are displayed in FIG. 15.

A weight loss of 16.5% w/w was observed in TGA associated with 2endothermic events in DSC recorded at 85° C. (onset) and 151° C. (onset)which could correspond to desolvation phenomena.

Example 6: Gravimetric Vapour Sorption (GVS)

Sorption isotherms were obtained using a SMS DVS Intrinsic moisturesorption analyser, controlled by DVS Intrinsic Control softwarev1.0.0.30. The sample temperature was maintained at 25° C. by theinstrument controls. The humidity was controlled by mixing streams ofdry and wet nitrogen, with a total flow rate of 200 ml/min The relativehumidity was measured by a calibrated Rotronic probe (dynamic range of1.0-100% RH), located near the sample. The weight change, (massrelaxation) of the sample as a function of % RH was constantly monitoredby the microbalance (accuracy±0.005 mg). Typically 5-20 mg of sample wasplaced in a tared mesh stainless steel basket under ambient conditions.The sample was loaded and unloaded at 40% RH and 25° C. (typical roomconditions). A moisture sorption isotherm was performed as outlinedbelow (2 scans giving 1 complete cycle). The standard isotherm wasperformed at 25° C. at 10% RH intervals over a 0-90% RH range. Dataanalysis was undertaken in Microsoft Excel using DVS Analysis Suitev6.0.0.7. The sample was recovered after completion of the isotherm andre-analysed by XRPD.

TABLE 1 Method Parameters for SMS DVS Intrinsic Experiments ParametersValues Adsorption - Scan 1 40-90 Desorption/Adsorption - Scan 2 90-0,0-40 Intervals (% RH) 10 Number of Scans 2 Flow rate (ml/min) 200Temperature (° C.) 25 Stability (° C./min) 0.2 Sorption Time (hours) 6hour time out

Form A

The mass change was <0.3% w/w between 0-90% RH. The material is nothygroscopic. No significant changes were observed in the XRPD after GVSanalysis.

Form B

The mass change was 2.3% w/w between 0-90% RH. No hysteresis wasobserved. No significant changes were observed in the XRPD after GVSanalysis.

Example 7: Thermodynamic Aqueous Solubility

Aqueous solubility was determined by suspending sufficient compound inwater to give a maximum final concentration of ≥10 mg/ml of the parentfree-form of the compound. The suspension was equilibrated at 25° C. for24 hours then the pH was measured. The suspension was then filteredthrough a glass fibre C filter. The filtrate was then diluted by anappropriate factor e.g. 101. Quantitation was by HPLC with reference toa standard solution of approximately 0.25 mg/ml in DMSO. Differentvolumes of the standard, diluted and undiluted sample solutions wereinjected. The solubility was calculated using the peak areas determinedby integration of the peak found at the same retention time as theprincipal peak in the standard injection.

TABLE 2 HPLC Method Parameters for Solubility Measurements Type ofmethod: Reverse phase with gradient elution Column: Phenomenex Luna, C18(2) 5 μm 50 × 4.6 mm Column Temperature (° C.): 25 Standard Injections(μl): 1, 2, 3, 5, 7, 10 Test Injections (μl): 1, 2, 3, 10, 20, 50Detection: 260, 80 Wavelength, Bandwidth (nm) Flow Rate (ml/min): 2Phase A: 0.1% TFA in water Phase B: 0.085% TFA in acetonitrile Time(min) % Phase A % Phase B Timetable: 0.0 95 5 1.0 80 20 2.3 5 95 3.3 595 3.5 95 5 4.4 95 5

Analysis was performed on an Agilent HP1100 series system equipped witha diode array detector and using ChemStation software vB.02.01-SR1.

The solubility of Form A in aqueous solution at different pH ispresented in Table 3.

TABLE 3 Form A Solubility in Aqueous Solution at Different pH pH Conc.Found (mg/mL) 1.2 1.3 1.64 1.07 1.95 0.82 3 0.10 4 0.022 5 0.017 6 0.0158 0.013 9 0.020 10 0.010

The thermodynamic aqueous solubility of Form B at pH of 7.42 wasdetermined to be 0.0096 mg/ml.

Example 8: Chemical Purity Determination

HPLC analysis performed on an Agilent HP1100/1200 system equipped with adiodearraydetectorandusingChemstationsoftwareusingthemethoddetailedbelow:

TABLE 4 Method Parameters Type of method: Reverse phase with gradientelution Test sample make-up: ~0.1 mg/ml in 60:40 v/v H₂O:ACN Column:Gemini-NX C18, 4.6 × 150 mm, 3 μm Column temperature (° C.): 40Injection (μl): 100 Detection: 260, scan from 210 to 500 nm Wavelength,Bandwidth (nm) Flow Rate (ml/min): 1.5 Phase A: 0.1% TFA in water PhaseB: 0.1% TFA in acetonitrile Time (min) % Phase A % Phase B Timetable: 075 25 10 70 30 30 65 35 45 35 65 46 10 90 50 10 90 51 75 25 60 75 25Retention time: ~20 min Needle wash: methanol:water (8:2)

In some embodiments, Form A is greater than 95% pure by HPLC analysis.In some embodiments, Form A is greater than 96% pure by HPLC analysis.In some embodiments, Form A is greater than 97% pure by HPLC analysis.In some embodiments, Form A is greater than 98% pure by HPLC analysis.In some embodiments, Form A is greater than 99% pure by HPLC analysis.In some embodiments, Form A is 99.8% pure by HPLC analysis.

In some embodiments, Form Bis greater than 95% pure by HPLC analysis. Insome embodiments, Form Bis greater than 96% pure by HPLC analysis. Insome embodiments, Form B is greater than 97% pure by HPLC analysis. Insome embodiments, Form Bis greater than 98% pure by HPLC analysis. Insome embodiments, Form Bis greater than 99% pure by HPLC B5207895.1-9analysis. In some embodiments, Form B is 97.8% pure by HPLC analysis. Insome embodiments, Form B is 99.8% pure by HPLC analysis.

In some embodiments, Form C is greater than 95% pure by HPLC analysis.In some embodiments, Form C is greater than 96% pure by HPLC analysis.In some embodiments, Form C is greater than 97% pure by HPLC analysis.In some embodiments, Form C is greater than 98% pure by HPLC analysis.In some embodiments, Form C is greater than 99% pure by HPLC analysis.In some embodiments, Form C is 99.4% pure by HPLC analysis.

Example 9: Chiral Purity Determination

Chiral purity of Compound 1 was determined by using a Lux Cellulose-1chiral column by normal phase HPLC. The mobile phase is composed of 20%isopropyl alcohol and 80% hexanes. The enantiomers of1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneare detected at 260 nm. In one embodiment, Compound 1 is dissolved in amixture of Hexanes: IPA=(7:3) to obtain a concentration of approximately0.2 mg/mL and the chiral purity of the sample is analyzed. The contentof the R enantiomer is determined by peak area normalization of theenantiomer peaks and is expressed in weight to weight percent. In someembodiments, a sample of Compound 1 includes less than 5.0%, less than4.0%, less than 3.0%, less than 2.0%, or less than 1.0%, of the(S)-isomer. In some embodiments, a sample of Compound 1 includes lessthan 1.0% of the (S)-isomer.

Solid Oral Dosage Forms

In some embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis formulated into a solid oral dosage form. In some embodiments, thecrystallinity of1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis maintained in the solid oral dosage form. In some embodiments,crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis formulated into tablets. In some embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis formulated into pills. In some embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis formulated into capsules. In some embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis placed into capsules without excipients or with excipients. In any ofthese embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis Form A. In any of these embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis Form B. In any of these embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis Form C. In any of these embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis Form D. In any of these embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis Form E. In any of these embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis Form F. In any of these embodiments, crystalline1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-oneis a mixture of two or more crystalline forms selected from the groupconsisting of Form A, Form B, Form C, Form D, Form E, and Form F.

Example 10: Capsule Formulations

In one embodiment, capsule formulations of Compound 1 for administrationto humans is prepared with the following ingredients:

TABLE 5 Capsule Formulations 40 mg Capsule 140 mg Capsule 140 mg Capsule200 mg Capsule w/w mg/ w/w mg/ w/w mg/ w/w mg/ Component % capsule %capsule % capsule % capsule crystalline 29.6 40.0 60.9 140.0 42.4 140.074.1 200.0 Compound 1 Microcrystalline 57.4 77.5 23.0 53.0 45.9 151.48.5 23.0 cellulose NF Croscarmellose 10.0 13.5 10.0 23.0 7.0 23.0 10.027.0 sodium NF Sodium lauryl 3.0 4.0 6.1 14.0 4.2 14.0 7.4 20.0 sulfateNF Magnesium NA NA NA NA 0.5 1.6 NA NA stearate NF

In some embodiments, the manufacturing process includes the followingsteps: weigh the indicated amount of the components, mix together andadd into an appropriate size capsule, and close capsule. In someembodiments, the capsules are stored at room temperature for an extendedperiod of time until they are used.

Example 11: Immediate Release Tablets

In some embodiments, tablets are prepared with the components set forthin Table 10.

TABLE 6 Components of Tablet Formulation Ingredient Range crystallineCompound 1 5% to 50% Hypromellose 2% to 10% Croscarmellose sodium 0% to15% Microcrystalline 5% to 50% cellulose Lactose 10% to 75% Magnesiumstearate 0.25% to 2.5% Total Tablet weight range: 300 mg to 1000 mg

Manufacturing process will typically be granulation (dry, wet or melt)or direct compression.

Example 12: Safety and Tolerability Study of Compound 1 in ChronicLymphocytic Leukemia

Purpose: The purpose of this study is to establish the safety andoptimal dose of orally administered Compound 1 (420 mg/day) in patientswith B-cell chronic lymphocytic leukemia/small lymphocyticlymphoma/diffuse well-differentiated lymphocytic lymphoma.

Primary Outcome Measures: Safety and tolerability of Compound 1(frequency, severity, and relatedness of adverse events).

Secondary Outcome Measures: Pharmacokinetic/Pharmacodynamic assessments.Tumor response—overall response rate as defined by recent guidelines onCLL and SLL (B cell lymphoma) and duration of response.

Eligibility: 18 Years and older; both genders are eligible.

Inclusion Criteria: 1. For treatment-naive group only: Men and women≥65years of age with confirmed diagnosis of CLL/SLL, who require treatmentper NCI or International Working Group guidelines 11-14. 2. Forrelapsed/refractory group only: Men and women≥18 years of age with aconfirmed diagnosis of relapsed/refractory CLL/SLL unresponsive totherapy (i.e., failed≥2 previous treatments for CLL/SLL and at least 1regimen had to have had a purine analog [eg, fludarabine] for subjectswith CLL). 3. Body weight≥40 kg. 4. ECOG performance status of ≤2. 5.Agreement to use contraception during the study and for 30 days afterthe last dose of study drug if sexually active and able to bearchildren. 6. Willing and able to participate in all required evaluationsand procedures in this study protocol including swallowing capsuleswithout difficulty. 7. Ability to understand the purpose and risks ofthe study and provide signed and dated informed consent andauthorization to use protected health information (in accordance withnational and local subject privacy regulations).

Exclusion Criteria: 1. A life-threatening illness, medical condition ororgan system dysfunction which, in the investigator's opinion, couldcompromise the subject's safety, interfere with the absorption ormetabolism of Compound 1 PO, or put the study outcomes at undue risk. 2.Any immunotherapy, chemotherapy, radiotherapy, or experimental therapywithin 4 weeks before first dose of study drug (corticosteroids fordisease-related symptoms allowed but require 1-week washout before studydrug administration). 3. Central nervous system (CNS) involvement bylymphoma. 4. Major surgery within 4 weeks before first dose of studydrug. 5. Creatinine>1.5× institutional upper limit of normal (ULN);total bilirubin >1.5×ULN (unless due to Gilbert's disease); andaspartate aminotransferase (AST) or alanine aminotransferase(ALT)>2.5×ULN unless disease related. 6. Concomitant use of medicinesknown to cause QT prolongation or torsades de pointes. 7. Significantscreening electrocardiogram (ECG) abnormalities including left bundlebranch block, 2nd degree AV block type II, 3rd degree block,bradycardia, and QTc>470 msec. 8. Lactating or pregnant.

Example 13: Safety and Efficacy of Compound 1 in Subjects WithRelapsed/Refractory Mantle Cell Lymphoma (MCL)

The primary objective of this trial is to evaluate the efficacy ofCompound 1 in relapsed/refractory subjects with Mantle Cell Lymphoma(MCL). The secondary objective is to evaluate the safety of a fixeddaily dosing regimen of Compound 1 (560 mg/day in the form of capsules)in this population.

Primary Outcome Measures: To measure the number of participants with aresponse to Compound 1.

Secondary Outcome Measures: To measure the number of participants withadverse events as a measure of safety and tolerability. To measurepharmacokinetics to assist in determining how the body responds to thestudy drug. Patient reported outcomes (to measure the number ofparticipants reported outcomes in determining the health related qualityof life).

Eligibility: 18 Years and older; both genders are eligible.

Inclusion Criteria: Men and women≥18 years of age. ECOG performancestatus of ≤2. Pathologically confirmed MCL, with documentation of eitheroverexpression of cyclin D1 or t(11;14), and measurable disease on crosssectional imaging that is ≥2 cm in the longest diameter and measurablein 2 perpendicular dimensions. Documented failure to achieve at leastpartial response (PR) with, or documented disease progression diseaseafter, the most recent treatment regimen. At least 1, but no more than5, prior treatment regimens for MCL (Note: Subjects having received ≥2cycles of prior treatment with bortezomib, either as a single agent oras part of a combination therapy regimen, will be considered to bebortezomib-exposed.). Willing and able to participate in all requiredevaluations and procedures in this study protocol including swallowingcapsules without difficulty. Ability to understand the purpose and risksof the study and provide signed and dated informed consent andauthorization to use protected health information (in accordance withnational and local subject privacy regulations).

Major exclusion criteria: Prior chemotherapy within 3 weeks,nitrosoureas within 6 weeks, therapeutic anticancer antibodies within 4weeks, radio- or toxin-immunoconjugates within 10 weeks, radiationtherapy within 3 weeks, or major surgery within 2 weeks of first dose ofstudy drug. Any life-threatening illness, medical condition or organsystem dysfunction which, in the investigator's opinion, couldcompromise the subject's safety, interfere with the absorption ormetabolism of Compound 1 capsules, or put the study outcomes at unduerisk. Clinically significant cardiovascular disease such as uncontrolledor symptomatic arrhythmias, congestive heart failure, or myocardialinfarction within 6 months of screening, or any Class 3 or 4 cardiacdisease as defined by the New York Heart Association FunctionalClassification. Malabsorption syndrome, disease significantly affectinggastrointestinal function, or resection of the stomach or small bowel orulcerative colitis, symptomatic inflammatory bowel disease, or partialor complete bowel obstruction. Any of the following laboratoryabnormalities: 1. Absolute neutrophil count (ANC)<750 cells/mm3(0.75×109/L) unless there is documented bone marrow involvement. 2.Platelet count<50,000 cells/mm3 (50×109/L) independent of transfusionsupport unless there is documented bone marrow involvement. 3. Serumaspartate transaminase (AST/SGOT) or alanine transaminase(ALT/SGPT)≥3.0× upper limit of normal (ULN). 4. Creatinine>2.0×ULN.

Example 14: Phase 2 Study of the Combination of Compound 1 and Rituximabin High-Risk Chronic Lymphocytic Leukemia and Small Lymphocytic LymphomaPatients

Purpose: The goal of this clinical research study is to learn ifCompound 1 combined with rituximab can help to control chroniclymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL). Thesafety of this combination will also be studied.

Rituximab (375 mg/m²) given intravenously (IV) on Day 1, Day 8, Day 15,and Day 22, then continued once every 4 weeks only on Days 1 duringcycles 2-6. Compound 1 started on Day 2 of cycle 1 at a dose of 420 mg(3×140-mg capsules) orally daily and will be continued daily.

Primary Outcome Measures: Progression free survival (PFS) [Time Frame: 3months]-progression free survival defined as the time interval fromtreatment to progressive disease or death, whichever happens earlier.Patients in complete remission (CR), partial remission (PR) or stabledisease (SD) are all counted as progression-free. Survival or times toprogression functions estimated using the Kaplan-Meier method.

Secondary Outcome Measures: Toxicity [Time Frame: 3 months]—toxicityreported by type, frequency and severity. Worst toxicity grades perpatient tabulated for selected adverse events and laboratorymeasurements. Toxicity (grade 3 or 4) monitored based on the Bayesianmodel (beta-binomial) by assuming a priori probability of toxicityfollowing beta(1,1).

Eligibility: 18 Years and older; both genders are eligible.

Inclusion Criteria: 1. Patients must have a diagnosis of high-riskCLL/SLL and be previously treated with up to 3 lines of prior therapy.High-risk CLL and high-risk SLL is defined by the presence of a 17pdeletion or 11q deletion or TP53 mutation. Any CLL and SLL patient whohas a short remission duration of less than 3 years after priorfirst-line chemo-immunotherapy, such as the FCR regimen, also fulfillscriteria of high-risk CLL/SLL, regardless of the presence or absence ofcytogenetic abnormalities. 2. CLL and SLL patients with 17p deletion orTP53 mutation will not be required to have received any prior therapy,given the poor outcome of CLL/SLL patients to standard frontlinechemo-immunotherapy, such patients will be eligible if they areuntreated or if they have received up to 3 lines of prior therapy. 3.Patients must have an indication for treatment by 2008 IWCLL Criteria.4. Patients age>18 years at the time of signing informed consent.Understand and voluntarily sign an informed consent. Be able to complywith study procedures and follow-up examinations. 5. ECOG/WHOperformance status of 0-1. 6. Patients of childbearing potential must bewilling to practice highly effective birth control (e.g., condoms,implants, injectables, combined oral contraceptives, some intrauterinedevices [IUDs], sexual abstinence, or sterilized partner) during thestudy and for 30 days after the last dose of study drug. Women ofchildbearing potential include any female who has experienced menarcheand who has not undergone successful surgical sterilization(hysterectomy, bilateral tubal ligation, or bilateral oophorectomy) oris not postmenopausal. Post menopause is defined as follows:Amenorrhea>/=12 consecutive months without another cause and adocumented serum follicle stimulating hormone (FSH) level>35 mIU/mL; amale of childbearing potential is any male that has not been surgicallysterilized. 7. Adequate renal and hepatic function as indicated by allof the following: Total bilirubin<=1.5× institutional Upper Limit ofNormal (ULN) except for patients with bilirubin elevation due toGilbert's disease who will be allowed to participate; an ALT</=2.5×ULN;and an estimated creatinine clearance (CrCl) of >30 mL/min, ascalculated by the Cockroft-Gault equation unless disease related. 8.Free of prior malignancies for 3 years with exception of currentlytreated basal cell, squamous cell carcinoma of the skin, or carcinoma insitu of the cervix or breast. 9. A urine pregnancy test (within 7 daysof Day 1) is required for women with childbearing potential

Exclusion Criteria: 1. Pregnant or breast-feeding females. 2. Treatmentincluding chemotherapy, chemo-immunotherapy, monoclonal antibodytherapy, radiotherapy, high-dose corticosteroid therapy (more than 60 mgPrednisone or equivalent daily), or immunotherapy within 21 days priorto enrollment or concurrent with this trial. 3. Investigational agentreceived within 30 days prior to the first dose of study drug or havepreviously taken Compound 1. If received any investigational agent priorto this time point, drug-related toxicities must have recovered to Grade1 or less prior to first dose of study drug. 4. Systemic fungal,bacterial, viral, or other infection not controlled (defined asexhibiting ongoing signs/symptoms related to the infection and withoutimprovement, despite appropriate antibiotics or other treatment). 5.Patients with uncontrolled Autoimmune Hemolytic Anemia (AIHA) orautoimmune thrombocytopenia (ITP). 6. Patients with severe hematopoieticinsufficiency, as defined by an absolute neutrophil count of less than500/micro-L and/or a platelet count of less than 30,000/micro-L at timeof screening for this protocol. 7. Any other severe concurrent disease,or have a history of serious organ dysfunction or disease involving theheart, kidney, liver or other organ system that may place the patient atundue risk to undergo therapy with Compound 1 and rituximab. 8.Significant cardiovascular disease such as uncontrolled or symptomaticarrhythmias, congestive heart failure, or myocardial infarction within 6months of screening, or any Class 3 or 4 cardiac disease as defined bythe New York Heart Association Functional Classification. 9. Significantscreening ECG abnormalities including left bundle branch block, 2nddegree AV block type II, 3rd degree block, bradycardia, and QTc>470msec. 10. Any serious medical condition, laboratory abnormality, orpsychiatric illness that places the subject at unacceptable risk ifhe/she were to participate in the study. 11. History of stroke orcerebral hemorrhage within 6 months. 12. Evidence of bleeding diathesisor coagulopathy. 13. Major surgical procedure, open biopsy, orsignificant traumatic injury within 28 days prior to Day 1, anticipationof need for major surgical procedure during the course of the study. 14.Minor surgical procedures, fine needle aspirations or core biopsieswithin 7 days prior to Day 1. Bone marrow aspiration and/or biopsy areallowed. 15. Serious, non-healing wound, ulcer, or bone fracture. 16.Treatment with Coumadin. Patients who recently received Coumadin must beoff Coumadin for at least 7 days prior to start of the study. 17. Anychemotherapy (e.g., bendamustine, cyclophosphamide, pentostatin, orfludarabine), immunotherapy (e.g., alemtuzumab, or ofatumumab), bonemarrow transplant, experimental therapy, or radiotherapy is prohibitedduring therapy on this study. 18. Use of medications known to prolongQTc interval or that may be associated with Torsades de Pointes (referto Appendix F) are prohibited within 7 days of starting study drug andduring study-drug treatment.

The examples and embodiments described herein are illustrative andvarious modifications or changes suggested to persons skilled in the artare to be included within this disclosure. As will be appreciated bythose skilled in the art, the specific components listed in the aboveexamples may be replaced with other functionally equivalent components,e.g., diluents, binders, lubricants, fillers, and the like.

The invention claimed is:
 1. A pharmaceutical composition comprising acrystalline form of ibrutinib and a pharmaceutically acceptableexcipient, wherein said crystalline form of ibrutinib is produced by aprocess, comprising combining a solvent solution comprising alcohol witha crystalline ibrutinib having 2-theta peaks at 7.0±0.1°, 14.0±0.1°,15.7±0.1°, and 18.2±0.1°; and a pharmaceutically acceptable excipient.2. The pharmaceutical composition of claim 1, wherein the alcohol isselected from the group consisting of: methanol and ethanol.
 3. Thepharmaceutical composition of claim 1, wherein the alcohol is ethanol.4. The pharmaceutical composition of claim 1, wherein the alcohol ismethanol.
 5. The pharmaceutical composition of claim 3, wherein thecrystalline form of ibrutinib has 2-theta peaks at 5.7±0.1°, 13.6±0.1°,16.1±0.1°, 18.9±0.1°, and 21.3±0.1°.
 6. The pharmaceutical compositionof claim 3, wherein the solvent solution further comprises water.
 7. Thepharmaceutical composition of claim 6, wherein the crystalline form ofibrutinib has 2-theta peaks at 5.2±0.1°, 10.2±0.1°, 16.5±0.1°,18.5±0.1°, and 20.8±0.1°.
 8. The pharmaceutical composition of claim 1,wherein the crystalline form of ibrutinib is recovered from the solventsolution.
 9. The pharmaceutical composition of claim 2, wherein thecrystalline form of ibrutinib is recovered from the solvent solution.10. The pharmaceutical composition of claim 3, wherein the crystallineform of ibrutinib is recovered from the solvent solution.
 11. Thepharmaceutical composition of claim 4, wherein the crystalline form ofibrutinib is recovered from the solvent solution.
 12. The pharmaceuticalcomposition of claim 5, wherein the crystalline form of ibrutinib isrecovered from the solvent solution.
 13. The pharmaceutical compositionof claim 6, wherein the crystalline form of ibrutinib is recovered fromthe solvent solution.
 14. The pharmaceutical composition of claim 7,wherein the crystalline form of ibrutinib is recovered from the solventsolution.